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Virus - Science topic
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Questions related to Virus
Hello all. I grew some cells for virus infection. I infected the cells with CMV (Cytomegalovirus) 8 days ago. The cells have not started lysis yet, but the plate is about to get dry. I wonder if I can add 5 mL of growth medium (DMEM) to the plate containing CMV-infected cells to prevent dryness at this moment (8 days after infection). Is there any risk to do so? Any advice is appreciated.
Hello all. I grew ARPE-19 cells in cell culture and infected them with a virus (Varicella-zoster virus). After the virus reached a high infection rate, I harvested everything in the plate and use freeze & thaw technique to release viruses into the supernatant. Now I want to store my viruses in -80 freezer. What is the composition of freezing medium for VZV? Are DMSO and FBS enough? Or do I need to add sucrose or something else? This is a bit urgent. I have no one to ask because I am the last employee in my lab as my professor is retiring. Any advice is appreciated.
I want to estimate the half-life value for the virus as a function of strain and concentration, and as a continuous function of temperature.
Could anybody tell me, how to calculate the half-life value in R programming?
I have attached a CSV file of the data
A number of definitions are found in publications ranging from 6 days to 14 days. Most frequent definitions mention 7 and 14 days. Some use onset of symptoms as staring point. Others use first detection of virus material as starting point. The end of shedding is usually defined by the last detection of viral material followed by negative sampling results. Usual duration of shedding (about 6 days) should probably be considered when the definition is chosen. Any suggestions?
Cancer as of now is not an infectious disease. The Papillomavirus, virtually, causes all cervical cancers and yet cervical cancer is "not considered" as an infectious disease? Some lung cancers of the smokers could, justifiably, be caused by papilloma virus and or other microorganismes, then why at least these cancers are not labeled as infectious diseases?
This question was extracted from the information from the journal “Tenants of Specimen Management in Diagnostic Microbiology” written by Rajeshwar Reddy Kasarla and Laxmi Pathak. The journal has relayed that most samples in Diagnostic Microbiology used for bacteriological examination or virus isolation were incubated, refrigerated, or incubated. Once the sample is received to be processed, what should be done to bring the sample back to its optimal temperature?
Does anyone knows the pH acceptable range for virus transport medium (VTM) for Sars cov 2 samples? I supose that it depends if you are only testing by PCR or if you need viability for culture but does anyone has experience in this subject?
Found a studie that defends that in normal individuals with no history of reflux or eustachian tube dysfunction, the pH values range from 6.10 to 7.92 with an average pH of 7.03 (SD, 0.67) so i believe that VTM should be buffered around pH 7 (with a variation of plus or minus 1) but need to confirm that.
Thank you and be safe.
Waiting for your suggestions.
Thanks
Uchurappa M
osmosis process occurs in bacterial and other cells.
But is it possible to virus?
Dear Researchers:
Could you please share some simple cures or prevention for COVID-19, Cold, Flu or Influenza, and possibly Other Viruses, and Cancers?
Updates on Oct. 10, 2023: First, many thanks to all contributors to this discussion. Here are some Natural Approaches found from surveying literature in medicine to Boost our Immune Systems against viruses such as COVID-19, Cold, Flu or Influenza infections and to avoid/minimize developing further inflammations in the lungs and hearts caused by some of those viruses:
Give it a try, please! Especially if you increase your Vitamin D level to a required level and consume Vitamin C sources, e.g., oranges, on a daily basis, you can check how rarely you would catch the virus. Or, even after catching the virus, the virus will likely develop very mild symptoms in your body.
1- Daily uptake of Vitamin D pills up to 100 IU per 1 kg weight is safe and very important, recommended by Afshar et al. (2000) and Dr. Hamid Sajjadi in an interview, to RAISE the Vitamin D level in our body to the POINT which is REQUIRED to BOOST our IMMUNE SYSTEMS against Viruses and Diseases including Cancers.
Vitamin D daily use needs to be adjusted based on our body weight.
Please read the following article by Afshar et al. (2000) about the importance of vitamin D and the required daily dose of it (Up to 100 IU per 1 kg weight) to boost our Immune Systems.
Please also read the following Review article by Jordan et al. (2022) about the importance of Vitamin D on the level of infection & disease progression for COVID-19. You may find in the article the importance of our Forgotten SUN.
Vitamin D is rarely available in food sources, except in fatty fish which needs to be eaten high enough to get the required amount of Vitamin D for a body.
Another good natural source is daily sunbathing with naked skin; however, in cloudy regions such as Europe, sunbathing doesn't work well.
Vitamin D helps to absorb Calcium in our intestines and thus, in order to avoid excessive absorption of Calcium by our body, it would be better to use Vitamin D pills with Calcium sources such as warmed-up milk and Magnesium sources such as bananas on a daily basis. Because magnesium competes with calcium in our intestines to get absorbed.
Here is a text from A Review article by Kulie et al. (2009) about some of the importance of Vitamin D on our health:
"Vitamin D is a fat-soluble vitamin that plays an important role in Bone Metabolism and seems to have some Anti-Inflammatory and Immune-Modulating properties. In addition, recent epidemiologic studies have observed relationships between low vitamin D levels and multiple disease states.
Low vitamin D levels are associated with increased overall and Cardiovascular mortality, Cancer incidence and mortality, and Autoimmune Diseases such as Multiple Sclerosis. Although it is well known that the combination of vitamin D and calcium is necessary to maintain Bone Density as people age, vitamin D may also be an independent risk factor for falls among the Elderly."
2- Having Good Nutrients including Protein sources, Minerals, and Other Vitamins, e.g., C, A, and E, sources from fresh fruits, vegetables, and nuts. For example, the good sources of fruits and vegetables for these vitamins could be a daily use of 1-2 Oranges for Vitamin C, Carrots for Vitamin A, and Almonds or Sunflower Seeds for Vitamin E.
As Vitamin C is a water-soluble vitamin, the excess of it will be excreted from the body, it needs to be consumed every day to provide everyday vitamin C requirements for the body, as it is the 2nd most important vitamin after Vitamin D to boost our Immune Systems against viruses and diseases.
And, Vitamin B family from grains, poultry, and meat sources.
3- After the infection by those viruses, gargling salty water to disinfect the throat to avoid further movement of the virus into the lungs as the virus may stay in the throat for a few days
4- Inhaling Steamed Fresh Leaves, if not available, the Oil, of Eucalyptus 4-5 times a day for several continuous days to kill the virus in the lungs.
Here is A Review article by Mieres-Castro et al. (2021) about the "Antiviral Activities of Eucalyptus Essential Oils: Their Effectiveness as Therapeutic Targets against Human Viruses"
Australian Aboriginals are very much using Eucalyptus to Treat Infections.
5- Having plenty of Warm Drinks to wash out the virus from our body and dilute the blood to avoid blood clotting.
6- Having enough sleep and daily activities/exercises
7- Kids are proven to have High Immunity Against COVID-19, likely due to having a high amount of Melatonin, the Sleep Hormone, in their blood. So, that is why kids sleep very much as you know.
Melatonin production in our body usually decreases with increasing age. Thus, we may use daily melatonin pills after the infection based on what physicians may prescribe for us.
Here is A Review article by Carrillo-Vico et al. (2013) about the Importance of Melatonin on the Functionality of Our Immune Systems:
8- Avoid Fear/Panic as it Substantially Deteriorates the Functionality of Immune Systems against viruses and diseases.
Here is an interview by Dr. Lauren Deville about How Fear Affects Our Immune System:
Hi, I've done a plaque reduction assay to analyze my possible plant activity, I used a HSV 2 virus at 6,5x10^3 as control, and I count (added file) the PFU... do I need to calculate something or I can make a graph with PFU results?
Akin to the oral "polio vaccine technology", is it possible for a healthy human to build antibodies against a said "bacterium, an example being a Steptococcal infection?".
In my opinion, yes we can. Our bodies have the required armoury "an adept immune system and the relevant enzymes", to tackle bacterial infections.
Please elucidate in detail, how you think our bodies could fight against a bacterial infection.
The previous discussion had been answered so well by "Rizzi". Looking forward to an answer like that of "Rizzi'.
I have been having an issue with my plaque assay for a few weeks now, I am working with a soil sample I inoculate a certain amount of virus PFU/mL into the soil then use some recovery method to recover the virus sample and run a plaque assay using methyl cellulose as an overly but the problem I keep having was mold growing on the sample which prevents seen any place on the sample. I would greatly appreciate any technical advice on how to get rid of the mold from the sample as we don't want to autoclave the soil or use UV radiation I would prefer using any mold inhibitory substance I have used Anti-Anti which it did not work as it only inhibits bacteria and fungi so is not efficient for my work. Thank you.
I am after the sequence for the murine leukemia virus-derived MND promoter (myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted), MNDU3. Can someone help me out?
Thanks Karin
I want to design tagman probe and primers to detect a virus. I had searched the complete DNA in NCBI.
What should I do next?
What is the standard procedure to design PCR probe and primers in general?
A patient with desminopathy (mutation Thr341Pro DES in a heterozygous state) with the progression of the disease has a decrease in taste and smell, immunosuppression, and an increase in IgA in the blood.
Oddly enough, but all this is characteristic of infections, including viral ones. For example, it is known that if the hepatitis C virus is not treated, then death will occur in 20 years.
In the identified case of late onset desminopathy, muscle weakness manifests itself at the age of 30, and death occurs 20 years after the onset of the disease.
Could the desmin mutation in myofibrillar myopathy be caused by an infection?
Perhaps the infection contributes to the progression of desminopathy?
Dear colleagues,
I defended my Ph.D. thesis in October 2016 and now I am looking for a postdoctoral position in microscopy (AFM, TEM, SEM) and biophysics of microorganisms (especially, viruses, I like them :)).
My CV is attached. If there is an open position in your lab, please, write me.
Best regards,
Denis
Various pandemic diseases have taught us various lessons from time to time, lastly, the spread of corona virus spread has shown how fickle human condition or survival is in face of sudden outbreak of dangerous diseases!
What are the human security implications of 'corona virus spread' around the world?
We would like to purchase around 10 thousand DNA oligos in a 96 well format (25 nmol). The cost per base is coming to around Rs 14-15. We wonder if there is any economical option available in the market.
Thank you
I have knocked down my gene of interest by lentiviral transduction. I revived the vials and continue growing cells for 72 h without any selection. After confluency, I passaged the cells later 24 hours I have used puromycin 1ug/ml, 3ug/ml, 5ug/ml for selection of transduced cells. After 2 days, most of my virus transduced cells die at all the puro conc. The viral transduction itself doesn't seem to be toxic as cells were growing up until selection. Any suggestions would be greatly appreciated!
I would like to increase the titre of my virus before spin infection to increase my infection efficiency.
The patient is being kept alive by 100% O2 input.
Q1
We have animal behavior scores of 4 group, Normal+ctrl virus, Normal+down-regulation virus, Model+ctrl virus and Model+down-regulation virus. It has two factors(Independent Variable): Model and virus. Editors suggested we use two way ANOVA to analyze, and now we obtained main effects of Model (F(1, 56)=201.18, P<0.0001) and virus (F(1, 56)=11.17, P=0.00427), as well as Model × virus interactions (F(1, 56)=16.13, P=0.0007).
If we should continue to calculate? For example, Model+ctrl virus vs. Model+down-regulation virus. We want to confirm the role of virus in Model animals.
Q2
Next, we used chemical drug to treat the Model animals and Normal animal. It has 4 drug concentration. Should we still use two way ANOVA to analyze the behavior scores? We want to know the role of different drug concentration in Model animals. And what do we do after two way ANOVA？
Thanks very very much!!!
Hi, here in my lab I have a kit for RT-PCR with probes (for the real time quantification). Can I run my reaction without adding the probes, in order to obtain en “end point” product? I was thinking about running two reactions, one without the probes and one with the probes, in order to obtain the product amplified but also its quantification.
I want to do this because I have to amplify a gene from a ssRNA+ virus and insert it into a plasmid.
Do you think it is possible? Does the absence of the probes impacts negatively on the Taqman polymerase? Thank you for your help
Physalis rugose mosaic virus (PhyRMV) is a Sobemovirus that causes severe damage to Physalis peruviana L., affecting vegetative parameters, fruit quantity and quality.
It was reported in Brasil and some other parts of South America. We are looking for photo of typical symptoms caused by this virus on host plants, especially on tomato.
I am trying to adapt a DNA extraction protocol to allow me to extract both RNA and DNA from the same sample. I plan to put the sample through Qiagen Allprep DNA/RNA mini kit, but I am not sure at what step I lose the RNA.
SDS/CTAB cleanup
a. Add 10 μl SDS (10%) + 1 μl proteinase K (10 mg/ml stock) to each tube and incubate at 56 °C for 20 min. At this point, pre-incubate CTAB/NaCl solution at 65 °C.
b. Add 35 μl NaCl (5 M) + 28.1 μl CTAB/NaCl (2.5%). Pulse vortex. Incubate at 65 °C for 10 min then perform a quick spin.
c. Add 200 μl phenol:chloroform:isoamyl alcohol (25:24:1) pH 8.0. Pulse vortex. Centrifuge 8,000 × g for 5 min at RT.
d. Collect the aqueous fraction. Add 200 μl chloroform. Pulse vortex for 3–5 sec. Centrifuge 8,000 × g for 5 min at RT.
e. Collect the aqueous fraction. This is final Virus Nucleic Acid.
The final viral nucleic acid goes through the Qiagen DNeasy Blood and Tissue kit.
I am not sure if the final nucleic acid contains RNA. If it does not contain RNA, I would like to know at which step I should put my sample through the kit.
I read protocols using CTAB to extract RNA as well as DNA, but I am not so sure about phenol:chloroform:isoamyl alcohol or plain chloroform. I read a similar protocol for extracting RNA that used CTAB and phenol:chloroform:isoamyl alcohol, but they replaced the chloroform with isopropanol and centrifuged, collecting the pellet as final RNA.
If someone could help me sort this out, it would be great!
FYI, this is part of a protocol to enrich for viral particles and extract the nucleic acid from stool with the least amount of human or bacterial contamination.
I am performing plaque assay from the spleen of mice infected with virus. I am facing a problem that in few mice i am getting plaques but in other mice samples i am not getting any plaque or less plaques in comparison to other mice of same group. I am not able identify the problem. I am washing spleen with pbs after collection and immediately storing this in liquid nitrogen. After this I am homogenizing spleen and performing plaque assay. Any help or suggestions will be appreciated. Thanking you in advance.
I've been having issues with viral production for the last couple of months. I was able to successfully make lentiviruses using a 2nd generation system 3 months prior and have a few aliquots frozen, but have unable to make more viruses. We recently tested the frozen nonconentrated virus by transducing a leukemia cell line and our transduction efficiency is about 80-90%.
My current protocol is:
Day -1) Seeding 293T on a 10 cm2 plate.
Day 0) Check to see if 293T confluence is 70-90%.
Prepare transfection complex:
Dilute packaging (psPax2), envelope (pRD114a) and transfer plasmid at 1:1:1 molar ratio in optimem.
Dilute PEI at 3X DNA MW in optimem.
Incubate for 10 mins.
Aliquot the PEI into the diluted DNA mixture. Incubate for 20 mins.
Add PEI:DNA mixture to 293T.
Day 1) Change media with fresh DMEM + FBS.
Day 2+3) Harvest supernatent. My transfection efficiency is about~80% evident by my GFP/mcherry reporter gene.
When I attempted to transduce the same leukemic cell lines, I was unable to detect my fluorescent reporter. It seems like even though my transfection was successful, my 293T are just not packaging the virus. Its not our transduction method because we are able to transduce with our old virus stock just fine. I tried different envelopes, different transfer plasmids, different aliquots of the packaging plasmids, freshly thawed 293T, different incubators, and different FBS manufacturers (both HI and non-HI). I could try different base medias incase the DMEM lot is bad, but i'm not sure if thats the case. I don't think it's out PEI because our transfection has been working well.
This dot appeard instead a visible distinct band. the concentration of the DNA was 30 microgeam/ml
Can you help me please if this is a band or not and why it appears like that ?
I am confused regarding protection of virus by wearing mask. The mask which is hurdle of inhaling of oxygen also. how we protect our selves from virus ?
I performed a transfection to obtain a recombinant virus that expresses my protein of interest, which is marked with a yellow fluorescent protein (YFP). The PCR results showed both the band of the expected size for the incorporated fragment and a smaller band that seems to be the parental virus with the YFP but without my protein of interest (because the ban's size is comparable to the lenght of the YFP's sequence). In this case I would have a mixture of 3 viruses: Parental virus, Parental virus+YFP and the virus with the YFP and my protein of interest. The last two both present the same fluorescence at the microscope so it's impossible to tell which one I'm selecting by clonal picking.
How can I separate the virus with the protein of interest from the one that only has the YFP?
I want convert the Titer of Virus which is given in 0.5ml to 1mL. tired many times but cant find a solution
I'm preparing virus sample for transmission microscopy for the first time. What is the best way how to concentrate virus for transmission microscopy? From literature, I understand that the best way is to use sucrose gradient, but I'll incubate virus with antibody as well. Is it safe to use ultracentrifugation for virus agregates or if will destroy the agregates? Or should I do ultracentrifuagtion first, then incubate with antibody and then do microscopy?
I have blood samples, and I want to detect a specific virus—whether it is present or not in these samples. I have extracted RNA and converted it to cDNA. Can Real-time PCR be used to detect the virus? I have a known sample for the virus as a refrence sample.
I am currently conducting research on RSV, and I am eager to know if the number of freeze-thaw cycles affects the titer after amplifying the RSV virus using HEp2 cells. Multiple freeze-thaw cycles may cause cell rupture and release of the virus, but repeated freezing and thawing of the virus can reduce the titer. Has anyone conducted relevant research and gained experience in this matter? Thank you very much.
As we observe the emergence of seasonal virus, with its peak transmission occurring during cold-and-flu season, what proactive public health measures should be implemented to effectively manage and mitigate the impact of this virus on public health systems, vulnerable populations, and overall community well-being?
I am having a doubt regarding CMV promoter and SV40 promoter in mammalian expression vector
1. Why CMV and SV40 promoters are used in mammalian expression vectors why not others?
2. There is a CMV enhancer and CMV promoter in some mammalian vectors why?
3. Why can't we use other virus promoters for driving transgene expression?
If you explanation for above questions kindly reply and mention any relevant articles.
It has been long ago when pandemic of HIV started.How does first person was affected of HIV virus through chimpanzees.
How can I analyze the lipid and protein content of a virus membrane or envelope? Are there any commercially available kits specifically designed to isolate the envelope from the virus, enabling further examination of the virus's lipid and protein composition?
Thanks.
I am been using VEROTMPRSS2 cells and incubating cells with 200ul of virus dilution for one hour. Cell layers are coming off after 30 min incubation. What might be the best solution to keep cell layer intact. is it with rocking the plate?
I am working with Rota virus and I am using MA104 cell line for propagation. I am not able to differentiate between mock flask and infected flask as it involves the addition of trypsin for both activation of virus and media used for the propagation. Does anyone have any more procedures related to the propagation of Rota virus and it's storage.
I had a miscommunication with a coworker, and as a result I put unfiltered virus harvested from 293T Phoenix media on to my primary fibroblasts. The transduction worked well, but now I have a couple contaminant Phoenix cells on my culture dishes.
I have tried several days of aggressive washing, since the Phoenix cells are easily detached, but that has been unsuccessful so far at removing all of them. The primary cells I have successfully transduced are very precious, so any other ideas at removing the Phoenix cells are appreciated.
As a last resort, I can flow sort the cells into single wells and grow them out from there. Hoping to avoid it for now as it is incredibly time consuming and also stresses out the cells.
People around the world are now nervous and confused about 'Corona Virus' . What are the differences between corona virus induced fever/cold/cough and normal fever/cold/cough ?
The high tide of coronavirus (COVID-19) has hit us again and again! And there are likely to be more deadly hits in the future. Meanwhile, preparations have been made to prevent the attack of a more deadly virus called 'Disease X'.
There is no way to stop this deadly virus. In this time of deep crisis, I have come up with two great ways to prevent viral infections including proper treatment of viral infections that can save countless lives. These methods will be effective in multiple ways. They are able to inactivate the virus and block and prevent the virus from entering the body cells. And the vaccine and the medicine I made will heal the person infected by the virus. And they are not at all harmful to our body.
To know more please watch video. Thank you
Hi all,
I am currently searching for articles about the structure of SRBSDV in tomo, but I have only found a few articles describing a part of the protein of this virus, and I have not found any articles about the overall structure of the virus in tomo, so do you have any clues?
Thanks a lot in advance.
Hello, I am working with closely related endogenous retrovirus (ERV) sequences. I suspect these might have been the result of several integration events from different but related exogenous retroviruses according to host phylogeny and geography. Basically, distant hosts that share geographical distribution have higher ERV identity between theirs than with closer species. ERVs from a single species that is closely related to a few others and has the same habitat, is however very different from its neighbors.
So two possibilities: each ERV clade comes from a different virus or all of them descend from the same insertion event.
But it remains as a hunch. Is there some sort of statistical test or other kind of test that might at least support or oppose this claim?
Thank you in advance.
For long Covid this might help. i just thought will send this. The acidic pH in absence of oxygen might help as the SARS-CoV2 virus had issue replicating in high altitude.
Those having long Covid *can do walking without inhaling air in breath.* To the level of comfort. In absence of oxygen the CO2 is built up. Say in cycles of one minute. This elevated CO2 influences the pH to acidic and that will interfere with the viral replication and provide immunity by altering & inactivating the virus. This absence of O2 will be anaerobic glycolysis with high lactic acid.
Acids have shown to denature the protein coat of virus. Here the CO2 formed in the body before being released from lung is being used to slightly alter the pH to inactivate the virus. The lactic acid also in muscles the way it stiffens in absence of oxygen.
When tested in positive long Covid patient One feels relief from symptoms after few days and the symptoms of long Covid reduce in intensity.
One can few times a day 3-4 say while walking for 5-10 minutes . Or once for say 30 minute walk what ever time one can exhale the breathe and continue walking. By understanding the level of one's comfort. THIS IS ONLY FOR PEOPLE WHO ARE NORMAL HEALTHY WITHOUT ANCILLARY CONDITIONS but have long Covid
Idea is using the acidic pH to alter the virus in the arterial blood by the CO2 forming carbonic acid and the acidic pH will do the work.
I can't find a search base for old variants of the SarsCov2 virus
At the GISAID database there are many areas besides the SEARCH too interface, where you can find this type of thing.
Hi,
I want to transfect my cell with two virus constructs requiring puromycin selection, but it already contains a construct with that selection. Should I add an extra amount of puromycin? The virus constructs contain RFP and GFP which I can sort on FACS, but even after that, I am afraid that the cells might kick out a gene or two. Any advice on what would be the best approach would be without tedious cloning?
Thank you
Restriction enzymes are common tools to compose genetically engineered plasmids in vitro. In laboratory circumstances it is possible to break DNA strands and recombine it with another strand.
However, I do not know whether such processes may happen spontaneously in cells that contain endonucleases and are co-infected with DNAs of different viruses? Or splitting and recombining DNA strands by endonucleases may only work with isolated endonucleases in laboratory settings?
Coronavirus vaccine.. What happens after the virus changes its configuration ? What will the live vaccine induce in our bodies? What will the immune response produce in our bodies ?
I'm working on microinjection. I wonder that how would- storage AAV2-Retro if it was not totally over in the pipette? Normally, we storage it in aliquots at -80. The virus I'm using now is left in the pipette. So, should I store it at -80 or -20? If it can be stored at +4, how long can it be stored? Finally, can I store AAV2-Retro with pipette?
Ernakulam Kerala kojijhor
Can you tell me affected nos of patients in this area and which action taken to prevent it.
Hi!
I need help with virus quantification and I am confused with the units. In some studies virus's titres are written as 10E5 TCID50/mL in other studies 5 log10 TCID50/mL. Is it the same?
Thank you
Hello, I've recently been studying Ancestral Sequence Reconstruction (ASR), attempting to infer ancestral sequences of viruses. I understand that this inference is constrained by factors like sample size and models, and represents a plausible sequence that may have existed. However, I'm curious about whether directly comparing these inferred ancestral sequences holds biological significance. Can they reflect the differences among the extant sequences from various lineages that were used to infer them?
Will the new mutated virus have the same effect as Corona virus as it did before?
Hello guy, I need your help to solve my problem in experiment. I got sample as culture cell line. I have to detect whether virus infection or not in this cell culture sample (pharmaceutics product). I should test the presence or not of RNA virus from bovine/porcine in these cell lines by quantitative real-time PCR method.
My plan is: I will find the specific sequence for these RNA viruses to design primer, probe and plasmid (that I will use as positive control when I running RT-qPCR, and make standard curve). In this case, when I perform RT-qPCR I need RNA or DNA to running.
My question here is: In this case, how can I isolate RNA or DNA of virus from culture cells? Or anyone have experience to perform this kind of experiment before can tell me what should I do in this case?
I have never do this kind of experiment before so I am so confused. Thank you so much for your help.
please mention the key differences and why best.
What type of proteins are those that make up virion particles? Same question is for ribosomal proteins.
Hello guy, I need your help to solve my problem in experiment. I got sample as culture cell line. I have to detect whether virus infection or not in this cell culture sample (pharmaceutics product). I should test the presence or not of RNA virus from bovine/porcine in these cell lines by quantitative real-time PCR method.
My plan is: I will find the specific sequence for these RNA viruses to design primer, probe and plasmid (that I will use as positive control when I running RT-qPCR, and make standard curve). In this case, when I perform RT-qPCR I need RNA or DNA to running.
My question here is: In this case, how can I isolate RNA or DNA of virus from culture cells? Or anyone have experience to perform this kind of experiment before can tell me what should I do in this case?
I have never do this kind of experiment before so I am so confused. Thank you so much for your help.
#SARSCoV2 is airborne - I am interested in finding the most recent / definitive research into its nature and protection from transmission. Thank you.
I know the word to be for human children but Hess writes in 1971:
"Others reported substantial attenuation after 100 passages in rabbits (MENDES, 1962). Another attenuated lapinized strain of ASF virus recovered it&initial virulence when passaged a number of times in pigs (SANCHEZ BoTIJA, 1962).
Russian investigators (KovALENKO et al., 1965) have shown that kids 4 to 5 months old could be infected with ASF virus by intraperitoneal inoculation of infected blood. The animals developed symptoms in 6 to 25 days and one kid died after 36 days. Virus was found in the blood 6 days after infection but was no longer present after 30 days. It was present in the spleen after 36 days but not after 70 days. The disease was characterized by hyperthermia, diarrhea, severe emaciation and by lesions in the reticuloendothelial system. The virus was passaged 19 times in kids and appeared to adapt progressively to these animals causing damage to the reticuloendothelial system and accumulating in the spleen [1]."
1. Hess, W.R. African Swine Fever Virus. Virol. Monogr. Virusforsch. Einzeldarst. 1971, 9, 1–33, doi:10.1007/978-3-7091-3987-5_1.
I searched through the internet but in vain. Also, I could not find the article of Kovalenko, either in English or Russian.
Thank you in advance
To date, the human cost of coronavirus (COVID-19) is more than 13 000000 infections, and more than 570000 death worldwide. The economic cost so far has been staggering. Many economies almost come to a halt. The impact on supply, demand, the financial market is affecting both larger and smaller firms. However, SMEs are at a disadvantage due to limited resources, existing obstacles in securing capital, and the span of time over which they can survive this pandemic compared to the larger firms.
How SMEs and new start-ups are going to handle this pandemic? Can they survive it or a great majority of them will go out of business? Should the government step in to help?
Since the start of the COVID-19 Pandemic, many governments and private organizations allocated large sums of money to fund projects dealing with various areas related to this virus. The vaccine is the most prominent area but detection, caring and monitoring of the patients revealed that the current medical equipment is not adequate and sufficient. Are these funding going to lead to invention or innovation? have you seen any report of innovation in medical technology in your community?
Hi,
I am introducing the strategy briefly which I follow to infect the MC38 cells with harvested lentivirus media from HEK293T cells at 24 and 48h.
1. First, MC38 cells are cultured in 60mm culture dish and when cells growth reach at 70% confluent, I change the previous media to virus medium for transduction with the help of polybrene (8ug/ml).
2. After that when cells growth is completed, I subculture the first-time infected cells and again transduction is done with viral media for second time when cells volume is 50-70%. Then the dishes are kept on incubator for overnight or more based on cells growth.
3. Finally, GFP positive cell percentages are determined by flow cytometry after proper growth. The problem is that GFP% is very low (<10%) although the GFP% of HEK293T cells are around 30%.
What should I do to increase the efficiency of infection?
Do somebody try to package virus >12KB plasmid? What is the efficiency?
Good day everyone, Please I am doing transfection/ transduction, I want to check the MOI of the virus and at the same time perform viral concentration, but i am stuck up with the steps. please can some assist me with relevant information ?
I am looking for a cell lineage infected by herpes simplex 1 virus.
PeiR is a lytic enzyme which is a methanogen virus that infects Methanobrevibacter ruminantium M1. Do all protease have active sites, if so how do I find it with the sequence only.
Hello everyone,
I am in the doing viral injection (hamilton and automatic pump) with AAV-mcherry targeting piramidal neurons in mouse aging from P40 to P50. After 2 to 3 weeks, I performed optogenetics stimulation, then after having the brain removed, soaked in formalin and fixated for at least 48h, I checked the viral expression with fluorescence microscope.
Here's the point: out of 10 trials, 4 times I didn't find any trace about the viral expression. I have checked the whole brain, but still zero. We are talking about zero level expression. On the other hand, in 6 experiements I have strong and proper expression.
In my experimental routine, I had 2 injections per day (2 mice). On the same days, I withdraw the virus out of the same small eppenderf tube each time for one animal. Between the two viral injections, the virus is kept soaked in ice in a dark water proof container.
I am sure the virus was injected (I saw the virus level in the glass capillary lowering). As always, I have waited 10 min before needle removal.
What could have happened in those 4 trials with no expression? Is the virus contained in the eppendorf damaged (at least in some of them)? I really don't know what to think right now.
Any help would be much appreciated.
I am writing a review article, I want to know if it is common to use a general 3d structure of a virus in the publication. (of course by mentioning the reference)
Which of the following is a characteristic symptoms of plants infected with phytoplasma that makes it different from a virus?
Can lentivirus vectors be directly transfected without packaging the virus？
In focus reduction neutralization assays, we apply a virus and serum mixture to a monolayer of cells. After infection, incubation, fixing, and staining we count the number of foci (formed from staining proteins released by dead cells) in order to determine if the sera applied to the cells blocked viral infection. We have been doing these assays for years and all of a sudden there has been a lot of variation in the virus phenotypes in our assays. We have a consistent incubator temperature, we have used the same cell line (vero-81 cells), same virus strains, media, etc. The only difference is different sera samples. Can anyone explain this?
Hello,
How would I determine the # of amino acid residues in a sequence? Is there an easier way to count them?
I'm trying to calculate the molar extinction coefficient for AAV. The MW of the AAV is 3740000 Da.
ϵ(280) (M−1 cm−1) = (#Trp)(5,500) + (#Tyr)(1,490) + (#cystine)(125).
Thank you!
Dear Researchers,
I need 2 ml of NZ2 strain ORFV virus for my student Ph thesis. Could you send us, buyer pay.
My address:
Prof.Dr. Mehmet KALE
Department of Virology
Faculty of Veterinary Medicine
Burdur Mehmet Akif Ersoy University
Burdur, Turkey.
E-mail: [email protected] or [email protected]
Any link will also be helpful.
Thank you
What is the likelihood of another pandemic in the future as estimated by the predictive analyses carried out, based on computerised, multi-faceted, big data mathematical modelling?
To what extent does climate change, progressive global warming, climate change across continents, increased environmental pollution and the impact of toxic waste pollution on human health, etc. increase the likelihood of another pandemic in the future as estimated by the predictive analyses carried out, based on computerised, multi-faceted, big data mathematical modelling?
On 4 May 2023, the World Health Organisation lifted the state of global epidemiological emergency associated with Covid-19. The WHO declared that Covid-19 no longer posed a public health, human health threat on a global scale. The WHO introduced the state on 30 January 2020, and after more than three years, the state was lifted. But the key point is that it was lifted as an epidemiological risk 'only' on a global scale and not as a direct recommendation for individual countries. Well, in individual countries, the levels of infection and mortality, although significantly lower than in 2020, are still occurring as part of local, successive, seasonal increases in infection with specific types of relentlessly emerging successive virus strains, and are significantly different in terms of the comparative analyses carried out. Globally, almost 7 million people have died according to Covid-19 death statistics and in more than 90 per cent of cases in combination with the presence of various co-morbidities. In Poland, these deaths were 120 000 with 5.5 million diagnosed infections and more than 250 000 excess deaths. In Poland, the Covid-19 epidemiological emergency is due to be lifted at the end of June 2023. In relation to this, is there still research being conducted by the WHO on the secondary effects of the Covid-19 pandemic? The 2018 Spanish flu was an avian flu that passed to humans. This was not the only such case in which a virus that causes disease in specific animal species started to infect and cause specific diseases in humans as well. It may have been similar with the SARS-CoV-2 (Covid-19) coronavirus, because before it started infecting humans it had previously developed in certain bat species, among others. It is likely that this virus acquired new features after the modification of its genome applied in laboratories, its effect was enhanced, it escaped from the laboratory and also started infecting humans. According to mathematical models of forecasting, which take into account population growth, increased population density in urban areas, low levels of sanitation in many parts of the world, low levels of availability of clean water in many economically poorer countries, the rate of creation of new strains of influenza viruses, coronaviruses, RSV, etc., which attack humans and certain animal species, the progressive process of global warming, climate change on different continents, increased environmental pollution and the impact of toxic waste pollution on human health, etc., it is likely that the virus will become more widespread in the future.
In view of the above, I address the following question to the esteemed community of scientists and researchers:
To what extent does climate change, the progressive process of global warming, climate change across continents, the increase in environmental pollution and the impact of toxic waste pollution on human health, etc., increase the probability of the appearance of another pandemic in the future as estimated by the predictive analyses carried out based on computerised, multifaceted, data-intensive mathematical modelling?
What is the likelihood of another future pandemic estimated from ongoing predictive analytical work based on computerised multi-faceted mathematical modelling with big data?
What is the likelihood of another pandemic occurring in the future?
What do you think about this topic?
What is your opinion on this subject?
Please respond,
I invite you all to discuss,
Thank you very much,
Best wishes,
Dariusz Prokopowicz
Regarding to the question (covid-19 is a natural virus or created by human) who we can realize the fact of this issue?
Reply or response should be supported by valid reference or reasoning.
Should there be vaccination in the face of a disease outbreak in a population where there are disease cases and obviously infected individuals?
WHO defines vaccination as, "Vaccination is a simple, safe, and effective way of protecting you against harmful diseases before you come into contact with them." (https://www.who.int/news-room/questions-and-answers/item/vaccines-and-immunization-what-is-vaccination).
In Chapter 4.18 of OIE - Terrestrial Animal Health Code - 10/08/2022, you can do a Ring vaccination around a herd of infected animals to contain the disease in animals susceptible to the disease (certainly still not infected).
In the book, "Trends in Emerging Viral Infections of Swines, Kyoung-Jin Yoon, Jeffrey J. Zimmerman, Antonio Morilla · 2008", it is stated on page 162 Section 5 on Classical Swine Fever Virus that, "Vaccination in Infected herds helps spread field virus". and also, "In endemically infected, vaccinated herds, there is selection for low-virulent CSFV strains".
Please share your views with references if any.
Hello.
Is there any option to design positive control other than using MEGA software? Does anyone know or expert using the MEGA Software?
Has any got experience packaging MSCV vectors with large cargo? Im in the final stages of plasmid design and so far the size is ~8.8kb.. I'm concerned the virus will fail packaging.
The highly-infectious disease is similar to Ebola, with symptoms including fever, muscle pains, diarrhea, vomiting, and, in some cases, death through extreme blood loss.
Hundreds of people have died from the virus in recent years, almost all in Africa.
According to the World Health Organization (WHO), on average, the Marburg virus kills half of the people it infects, with previous outbreaks killing between 24% and 88% of patients.
The virus was first identified in 1967 after 31 people were infected and seven died in simultaneous outbreaks in Marburg and Frankfurt in Germany and Belgrade in Serbia.
The outbreak was traced to African green monkeys imported from Uganda.
But the virus has since been linked to other animals.
I'm trying to simulate virus and nanoparticle interaction. I wonder what software can run this types of simulation? can I use VMD ??
Covid news – live: Cases soar again in India as doctors warn of ‘new symptom’
A new coronavirus strain dubbed Arcturus appears to be driving a surge in Covid-19 cases in India, prompting the country to resume vaccine production and sparking fears it could lead to a rise in cases in the UK and elsewhere.
India on Friday recorded 11,109 new Covid infections, the biggest jump in almost a year. The country’s active case count is now up to 49,662.
The XBB.1.16 strain, a sub-variant of Omicron, has been found in 22 countries, including Singapore, Australia, the UK and the US. Research indicates Arcturus could be one 1.2 times more infectious than the last major sub-variant, making it likely to become the dominant strain.
The spread of the strain, first detected in late January in India, is worrying experts, as it seems to exhibit unique symptoms in children, one of which is conjunctivitis.
The symptoms of the variant include high fever, cough, and “itchy” conjunctivitis or pinkeye, according to Vipin Vashishtha, a paediatrician and former head of the Indian Academy of Pediatrics Committee on Immunisation.
COVID-19 and Influenza Activity
April 2, 2023 to April 8, 2023
These images provide a high-level assessment of respiratory virus activity in Ontario. Provincial percent positivity can be used to provide an estimate of the intensity of circulating viruses in the province. Percent positivity for the most recent week is used to assign influenza and COVID-19 to either a low, moderate, high or very high category. Weekly indicator change was determined by considering a combination of indicators (see Technical Notes). For further details, please refer to the Respiratory Virus Overview in Ontario report.
Other viruses are routinely isolated, but not SARS-CoV-2.
HeLa cells were resistant to TGEV infection. However, when I incubate the Hela cells with about 1 MOI of TGEV, a lot of cells died two days after incubation. Why does this happen? is it due to the stress from this virus?
Hello Everyone
I have been working on 14 different virus strains to be detected utilizing TaqMan Probes. Although I get good results in Singleplex assays with the almost same Ct for all samples, in multiplex assays 3 types have higher CTs (more than 5 cycles), Do u have any suggestions to overcome this issue? the plasmid copy number, primer-probe concentration, and qPCR probe Master mix are the same.
There are several health and physical problems, after recovering from the Corona virus, and I need research links on this topic, thank you very much for all
I am conducting Avian Influenza virus detection by real time rt PCR.
I did 4 target AI genes and 4 primer probe sets for PCR.
(H7 HA, H5 HA, NP and M target gene)
I run real time 4 time individually for that 4 target genes.
The result came out non-similarity.
(For Example,
In Sample no 1, H7 HA and H5 HA amplified (positive) but not amplified in NP gene and M gene .
In Sample no.2., result came all 4 gene were amplified.
In Sample no.3 NP gene and M gene and H5 HA gene came out positive result.)
How can I understand the nature of antigenic protein surfaces of virus and detection system of real time PCR.
Even though the same sample and same virus, can different results by using different target gene?
Hello,
I have a question regarding qPCR efficiency calculation.
Is it correct to talk about PCR efficiency if I perform a serial dilution of sample that containing an unkwon concentration of the viral particles?
I usually read that the serial dilution should be done from a known concentration of the extracted DNA. But I have stool samples where the amount of virus is unknown and the dilution was made from the sample stock. Afterwards the DNA was extracted and PCR was run.
When I calculate the slope of standard and the efficiency. Can I then talk about PCR efficiency?
Thank you for your feedback!
We are testing virus titer, and have to dilute the virus stock at 1:10,000 (Dilution A) and 1:20,000 (Dilution B) to run qPCR as templates. Then convert the diluted titer to dilution corrected titer. However, the conversion from A or B is not the same value, showing a big difference. I guess the problem is about dilution. Is there any best strategy procedure to complete the dilution within 3-step? Thank you.
During the thawing of the subpolar permafrost, triggered by accelerating global warming, could viruses and bacteria from many thousands of years ago, which are dangerous to humans, emerge and cause another pandemic?
The thawing of permafrost, which has been present for thousands and millions of years in areas near the Arctic Circle, mainly in the Arctic, caused by the accelerating process of global warming, will result in the release into the atmosphere of thousands and possibly millions of tonnes of hitherto frozen methane, a gas that is many times more greenhouse-generating than CO2, which will result in a significant acceleration of the already rapid process of global warming. However, this is not the only very dangerous effect for human civilisation and for the state of the planet's biosphere of the progressing process of global warming, a process which has been taking place since the first industrial revolution, i.e. since the 18th century. Among the significant negative consequences of the increasingly rapid global warming process triggered by the industrial revolution based on the dirty energy of burning fossil fuels is the increase in the risk of a future pandemic caused by viruses emerging from the thawing of the permafrost in areas near the planet's Arctic Circle. These viruses emerged and were frozen many thousands and perhaps millions of years ago, i.e. when there was not yet a modern species of homo sapiens on planet Earth. Therefore, humans may not be immune at all to these strains of different types of viruses that functioned on the planet many thousands of years ago. In addition, the existence of many species of both wild animals and farmed livestock may also be threatened if thawing viruses from many thousands of years ago prove to be completely unfamiliar to the immune systems of said animals. According to CNN media reports, there are virological research laboratories currently working on revived viruses taken from thawing permafrost. These revived viruses are referred to in the media as "zombie viruses". In addition, high summer temperatures have thawed the corpses of people who died and were buried in cemeteries many years ago, as well as animals, from whose thawing bodies pathogenic strains of viruses and bacteria have emerged. The thawing of the permafrost in recent years, for example, has been identified as a major source factor in the occurrence of the anthrax epidemic in Siberia, because the high temperatures experienced in Siberia for the first time in many thousands of years allow viruses and bacteria to be released from human cemeteries and animal corpses, i.e. micro-organisms that functioned thousands of years ago and which may be particularly dangerous to humans and animals living on the planet today.
In view of the above, I address the following question to the esteemed community of scientists and researchers:
In the course of the rapid thawing of the sub-polar permafrost, caused by the progressive process of global warming, could viruses and bacteria from many thousands of years ago, which are dangerous to humans, come to light and cause another pandemic?
What is your opinion on this subject?
Please respond,
I invite you all to discuss,
Thank you very much,
Best regards,
Dariusz Prokopowicz
Hello,
My topic of research is Avian Leukosis Virus (ALV). I have noticed that there are some old research papers that showed that when injecting chicken cells infected with Rous Sarcoma Virus (a close relative) in the brain, tumors would appear in non-avian animals such as mouse or monkeys. And those tumors are of host origin.
Basically they conclude that a virus specific to birds, that has not been reported to infect non-avians, can cause disease if introduced in the brain through experimental methods.
I haven't found current papers on this issue so my question is: Do we know of a mechanism that explains this? or is the methodology flawed? Do you know of a similar phenomenon in other viruses?
I harvested a virus. When I did plaque assay to titer it, I did duplicate and the titer was different. How can I take one of them? Will I average them? The concentrations were 4.50E+05 and 2.50E+05 PFU/ml.
I will do a PRNT test with these virus. Therefore, It is important to know its titer as accurately as possible! Any help would be greatly appreciated!
I have two DEG sets for 2 disease conditions (from mild to severe condition) of the same viral infection. When I look at the common gene from these two sets of DEGs, I found that some genes show opposite expression among these two conditions ( Like a gene downregulated in mild but up-regulated in severe or vice versa). So what I want to know is that,
1) If this phenomenon is normal in viral infection??
One of the most prevalent emerging pathogen that has turned into a public health issue in the recent years until now, is the virus that caused COVID-19 pandemic, known as the SARS-CoV-2. How may this virus potentially affect blood donations, and what measures may be taken by the blood bank to ensure the recipients' safety?
Which viral infection might be more severe?a transported virus from an animal to a human or a virus directly invade human?
