HCV RNA detection by TMA ?

[james744]

after exposure to check ones infection status , elisha need to be done at 6 months , pcr 6 weeks , I have heard TMA is more sensitive than pcr can i do it in 3 weeks , to make sure that i am ok ?

[james744]

When i Called the lab and asked about the method that they use for PCR they said that ...
they mailed me this ...
"For the HCV qualitative assay viral RNA is isolated from serum/plasma. For Reverse Transcription and PCR amplification, nested PCR is being employed. In this PCR system 2 sets of primers-outer set targeting bigger fragment of viral genome (5'NCR) and the inner set complimentary to regions inside the bigger fragments are used for amplification. Thus the amplified product of first step acts as a template to the nested step or second step. Detection of amplified sequence is done by 2% agarose gel electrophoresis."

they said RT-PCR .. is it sensitive enough to get tested at 3rd week ?

Sorry this may be a stupid question . But as it costs more , I am concerned about the sensitivity of the test at 3rd week and reliability of the test . So that i could have peace of mind after the test .

[Bill1954]

You should discuss shipment methods and test parameters with the lab tomorrow. Good luck with the results, James--

Bill

[james744]

Thanks BILL and Trinity

Tomorrow i am going to call the lab and going to ask about the sensitivity of the tests done . And will be sending the blood samples ( PACKED IN DRY ICE, Will it be enough guys ????) . It will take a day or 2 to for a sample to reach them .

And BILL is HCV bDNA suit me ? Its just 17 days post exposure ?

[Bill1954]

The sensitivity of HCV testing is critical in those that are undergoing antiviral therapy, James; and any certified clinical laboratory should be adequate for your needs.

The sensitivity is less of an issue in a patient that is not in antiviral treatment; the viral load for those patients would typically be in the 100.000 or 1.000.000 range.

In fact, a different, older assay that might be appropriate for you is the HCV bDNA qualitative assay, with a sensitivity of <615 IU/mL. This is less expensive and probably suitable for your current needs.

Good luck,

--Bill

[james744]

Thanks . I hope results will come good .

And i am from india. Here it is not much developed as US, so i am worried about the sensitivity of the machines over here .

[james744]

Thanks . I hope will come good .

And i am from india. Here it is not much developed as US so i am worried about the sensitivity of the machines over here .

[james744]

I dont know what is the sensitivity of the qualitative assay used .
I am form Indai, Labcorp Test centers are not available here . I am planning to give it to the ranbaxy labs .

I am not sure about the sensitivity of the assay used for qualitative analysis . But for the quantative analaysis the sensitivity is given as 15 IU/ml .

Will it be enough at 3rd week ? and will sensitivity of the qualitative test be more than this???

I tried to find the sensitivity of the qualitative test done here but its not specified .only quantitative is specified as 15 IU/ml

[Trinity4]

LabCorp Test

Hepatitis C Virus (HCV), NGI UltraQual™
Synonyms:
HCV UltraQual™
NGI UltraQual™
UltraQual™ HCV

Test Number: 140609 CPT Code: 87521  
Test Includes:  Detection and confirmation of hepatitis C virus


Use:  Detect and confirm hepatitis C virus infection

Limitations:  Assay sensitivity: 5-8 copies/mL (2-3 IU/mL)

[Trinity4]

HCV RNA by PCR can detect the presence of viremia after two weeks of possible exposure.  There are very sensitive tests which can go down to <2iu/mL.  A qualitative test would be cheaper and just as effective because it will detect the presence of HCV and if you were infected you would be in the acute state so viral load at this point would not be an issue.

As previously stated, your risk of becoming infected is slight.  Have the test and put you mind to rest.  Retest in 6 months.....done deal.

Trinity

[james744]

No Bill i am struck in home . Form the knife which had cut  my wife who has hep c + . we both are hep a and hep b vacinnated and hiv -ve.

By mistake she has not bleached it and she is more stressed and feeling pain more than me in this.

But it was rinsed well in running water. I touched it after 10 hours .
Will there be decrease in load of virus when it is outside the body ? or else it will start to decrease after 16 hours ?

[Bill1954]

Hi again, James—

HCV is typically an asymptomatic disease; most of us are unaware we’re infected until our doctor advises us of test results. I’m unaware of bone of joint pain being listed as a symptom of acute infection.

To my knowledge, all genotypes are equally detectable. If HIV coinfection is present, it can delay seroconversion (HCV antibody production), delaying HCV diagnosis. If you had a needlestick exposure, it’s advisable to test for other pathogens as well. I imagine you’re employed in health care? If so, this should be reported to a supervisor so that appropriate Post Exposure Prophylaxis (PEP).

You can read more here:

http://www.cdc.gov/hepatitis/Settings/HealthcareSettings.htm

--Bill

[james744]

another confusion due to way of my exposure to contaminated knife .

All site are saying HCV will be on surface for atleast 16 hours .  Will there be decrease in viral load as soon as it is exposed to environment ?

[james744]

Can all genotypes found in 2-3 weeks or is there any exceptions for genotype which takes more time to replicate ?

[james744]

Thanks For your reply and help BILL .

Actually i am very much worried and confused too . It has been 17 days since the last exposure . I have posted the exposure story here . I was like accidental needle stick exposure.

Can you please clear me one another confusion please ........

Some sites are saying to get tested after 6 weeks and some sites says PCR can detect in 2-3 weeks after infection . What shall i do ?

and I dont have fever or anyother illness now , but i feel little pain in one of my legs . is leg pain also the symptom of acute infection ?

[Bill1954]

This is from the U.S. Center for Disease Control and Prevention:

http://www.cdc.gov/hepatitis/HCV/HCVfaq.htm#section3
~~~~~~~~~~~~~~~~~~
Q) How soon after exposure to HCV can anti-HCV be detected?
A) HCV infection can be detected by anti-HCV screening tests (enzyme immunoassay) 4–10 weeks after infection. Anti-HCV can be detected in >97% of persons by 6 months after exposure.
Q) How soon after exposure to HCV can HCV RNA be detected by PCR?
A) HCV RNA appears in blood and can be detected as early as 2–3 weeks after infection.
~~~~~~~~~~~~~~

I’d think either TMA or PCR methodology would be appropriate for this test. There is however, a chance of intermittent viremia in the early stages of infection (acute infection). It’s still a good idea to check again at six months to rule this scenario out.

Good luck—

--Bill