exposed to lentivirus

[chanel_lee39]

Hi fellows,

 

This sounds really stupid but I spilled some lentiviral proteins onto my face today when I was loading the proteins in the SDS-PAGE gel. I am not sure if they got into my eyes but I wiped them out immediately after spilling. And the amount was very small. The proteins were collected from 293 cells that were used to package lentivirus so they definitely have lentiviral particles in them. I lyzed them with NP40 lysis buffer, they were sonicated and of course denatured by boiling. Were these procedures enough to kill all the virus or am I still at risk of getting infected? I am just worried that is there any chance the viral particles could revive once they get in vivo?

 

Thanks!

Chanel

Edited September 13, 2013 by chanel_lee39

[Endy0816]

You really need to report it to a supervisor. Your lab or facility should have some kind of procedures in place.

 

Without knowing the different variables there is no way to determine your risk.

 

 

Pathogenicity: Hazards depend on multiple factors: whether the vector is capable of infecting human cells, whether the vector is replication competent, how many viral genes are contained in the vector, and the specific transgenes present in the vector. On an emergent basis, the PI should be the best source of information regarding potential health hazards.

Modes of Transmission: Virus may be transmitted in the following ways: 1) a skin puncture or injection, 2) ingestion, 3) contact with mucous membranes (eyes, nose, or mouth), 4) contact with non-intact skin, and 5) low risk exposures include bites from an animal inoculated with lentivirus, percutaneous contact with body fluids from an animal inoculated with lentivirus and aerosols.

Incubation Period: Variable, may be months to years

Communicability: Replication incompetent vectors: Not communicable.

[chanel_lee39]

Thank you. I will talk to the P.I. The viral vector is pLVX-Puro from clontech and it does not have any insert in it. I am not sure if any got into my eyes. And the samples were processed through SDS lysis, sonicated and denatured at 95 Celsius. They were in sample buffer when I splashed them, only a tiny bit got out onto my face. Is there a chance the virus could reform in this case?

[CharonY]

Based on the description the actual risk appears to be low. Nonetheless, incidents like this have to be reported and documented for your own safety as according to biosafety regulations (in most countries, I presume).

That being said it should be noted that it sounds to me that several safe regulations are not being followed in your lab. The first is that every member of a lab (regardless whether student or staff) has to be informed on all potential biosafety hazards as well as risk assessment and safety procedure. I am aware this is sometimes done in a lax way, but considering that you have not immediately notified the incident appears that you have not been well informed. This is not to admonish you, but just to give you basic information on lab safety issues.

On the same note, all work with lentiviruses (as well as human cell lines for that matter) are automatically biosafety level 2, which also means that personal protection has to be used at all time. Technically this is also true for biosafety level 1 but people usually start to get really serious at 2.

So while doing work that can generate splashes or aerosol face protection (goggles) have to be worn. For your own protection, review the safety procedures, even if others may not follow them. In the end everyone is responsible for his/her own safety, even if the PI make think differently.

Edited September 13, 2013 by CharonY

[Endy0816]

Here is the manufacturer safety information:

http://www.clontech.com/xxclt_ibcGetAttachment.jsp?cItemId=42386&minisite=10020&secItmId=15021

 

CharonY's assessment of the situation is exactly right though.

[chanel_lee39]

Thank you all for the information. Yeah, my lab does have a lack of safety protection or education about this kind of stuff. My PI thinks that if everything is denatured, I should be safe. But I am just wondering can the virus somehow renature after thermo and detergent denaturation once they get in the body? Anybody knows?

[CharonY]

Considering the nature of the loading buffer and the heat denaturation plus further dilution after coming in contact with eye fluids risks of actually having intact particles are very low.

But even so I would always ask students (especially beginners) to treat everything as highly infectious. Simply as part of the training. You will get sloppier at some point anyway, may as well start out with a higher standard.

[chanel_lee39]

Thank you CharonY. Couldn't agree with you more. I have learned my lesson this time. I think more care really needs to be taken on this matter in our school.