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Itsminor

Affordable PCR Machine - swab yourself?

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Sanguine108

aw that's an adorable lil' thermocycler!

If you have the money then sure, go for it. If it's going to be a financial strain that could be better used elsewhere then save it.

Make sure you store the reagents and enzyme properly.  Next to the thermocycler, the enzyme would be the next expensive item.  ...But by only sampling yourself I don't think it'd be that bad.

...Makes me wonder about false positives though. PCR will amplify any small tidbit of target DNA, whether it's live or inactive.   This could be a problem as HSV is inactive without its lipid envelope, so HSV DNA can be around but 'dead'.  So if you do this, also swab a 'clean' area several inches away from suspected spot.  Best you can do as a control.

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Itsminor
1 hour ago, Sanguine108 said:

aw that's an adorable lil' thermocycler!

If you have the money then sure, go for it. If it's going to be a financial strain that could be better used elsewhere then save it.

Make sure you store the reagents and enzyme properly.  Next to the thermocycler, the enzyme would be the next expensive item.  ...But by only sampling yourself I don't think it'd be that bad.

...Makes me wonder about false positives though. PCR will amplify any small tidbit of target DNA, whether it's live or inactive.   This could be a problem as HSV is inactive without its lipid envelope, so HSV DNA can be around but 'dead'.  So if you do this, also swab a 'clean' area several inches away from suspected spot.  Best you can do as a control.

Amazing thanks! So you know about this area it seems! What do you mean by the enzyme?

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Sanguine108

so it's probably best to watch a youtube video but there's a few ingredients that go into the test tube.  

-your sample of interest that may or may not have HSV DNA in it.
-the test reagent that has different ions for the reactions and nucleotides for amplifying the sample of interest
-primers for target HSV
-the enzyme that is going to do all the work... Taq polymerase.

 

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WilsoInAus
9 hours ago, Itsminor said:

Check this out..

http://www.minipcr.com/?gclid=CMneifevvdQCFRQUGwodvgALsw

Could we use it to swab ourselves every day?

I'd personally like to know how often I'm shedding and it'd be handy to know if that red dot was actually herpes - or just a spot!

 

 

The trouble is though that it is probably shedding that happens during sexual intercourse that is the most likely to result in transmission.

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Itsminor
16 hours ago, WilsoInAus said:

The trouble is though that it is probably shedding that happens during sexual intercourse that is the most likely to result in transmission.

Oh is that so? I thought maybe I'd transmit while having a coffee in starbucks :D ahaha 

Yes, but I'd love to know how contagious I am in general...

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Itsminor
On 2017-6-14 at 2:40 PM, Sanguine108 said:

aw that's an adorable lil' thermocycler!

If you have the money then sure, go for it. If it's going to be a financial strain that could be better used elsewhere then save it.

Make sure you store the reagents and enzyme properly.  Next to the thermocycler, the enzyme would be the next expensive item.  ...But by only sampling yourself I don't think it'd be that bad.

...Makes me wonder about false positives though. PCR will amplify any small tidbit of target DNA, whether it's live or inactive.   This could be a problem as HSV is inactive without its lipid envelope, so HSV DNA can be around but 'dead'.  So if you do this, also swab a 'clean' area several inches away from suspected spot.  Best you can do as a control.

..Makes me wonder about false positives though. PCR will amplify any small tidbit of target DNA, whether it's live or inactive. 

Would this explain higher PCR shedding rates than culture? 

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Sanguine108
On 6/15/2017 at 11:11 AM, Itsminor said:

..Makes me wonder about false positives though. PCR will amplify any small tidbit of target DNA, whether it's live or inactive. 

Would this explain higher PCR shedding rates than culture? 

Yeah, I'm not very familiar with how viruses are processed for PCR.  I can't imagine it being different from animal or bacterial cells.  The primers for PCR shouldn't be able to bind/amplify/recognize the viral DNA within its protective capsid/shell so some sort of lysing agent would be necessary to dissolve it. (at least that's what goes on with animal and bacterial cell testing... to lyse the cell walls and liberate the DNA)  So then there's the question of environmental nucleases that degrade DNA.  Is there something that hinders the capsid's integrity, thereby allowing the HSV DNA out and vulnerable to nucleases that will degrade it?  If this were the case then what's the life-span of an HSV virus (no lipid envelope) capsid in the open environment?  I'm sure these questions have been answered but I don't have a reason to know them.

So to answer your question, I don't know. :)   If someone does know the answer then please share.

 

Edited by Sanguine108

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dont quit!17
15 minutes ago, Itsminor said:

Probably easier just to leave it and get my life back ahahah

Is the whole kit $4,500.00? I read some of the link and seen several kits but unsure which one is needed. This pretty interesting especially since Gen-003 will probably get approved. Some of the trial members from Genocea have decreased shedding to 0%. 

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Itsminor
21 hours ago, dont quit!17 said:

Some of the trial members from Genocea have decreased shedding to 0%. 

Really? 

Man it's crazy how much hope I'm putting on this thing. CAN YOU IMAGINE?!

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dont quit!17
2 hours ago, Itsminor said:

Really? 

Man it's crazy how much hope I'm putting on this thing. CAN YOU IMAGINE?!

Yeah and well others were above the average shedding rate. But this kit would be interesting to have nonetheless. 

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