PCR sensitivity test help

[Macola]

Hello again,
Today I went to the Institute where PCR tests are done, asking for some answers regarding their sensitivity results.
They told me that they measured from 400 - 6 000 000 copies (the results are measured in copies), so I wanted to know if it is sensitive enough to detect HBV. They also said that they could detect even one copy if it existed, so I am a little bit confused. My test result reads:

METHOD: AMPLICOR HBV Monitor Kit, Roche Molecular Systems, USA;
                PCR amplification of Hepatitis B virus

RESULT: The results from the serum analysis of the patient showed ABSENCE of Hepatitis B virus.

Please someone answer if they are sensitive enough, because I am also HBsAg negative, but for 3 years now I still don't have Anti HBs (I had an acute Hepatitis B). Should I make a biopsy or something to see for sure that there's no virus? Or this tests are enough sensitive?

[Macola]

Grateful for you advice.

I'll send blood samples to Synlab GmbH & Co. KG, Germany to do the pcr (I found a lab that cooperates with them and they send blood samples there for 120 euros).

Sorry for bothering you, and thank you for answering all of my questions. You are the best doctor I've known.

Best wishes and health

[stef2011]

architect is the best for quantification and also gets almost all hbsag mutants.it gets even a little lower than 0.05iu/ml but when hbsab i not present in the blood there is usually very low level hbsag in the liver

check if they can reach about 6iu/ml for hbvdna, if hbsag is <0.05iu/ml and hbvdna<6iu/ml chances of occult hbv making damage should be little and making serial monitoring every 3-6 months you should be able to detect some hbvdna sooner or later.

architect can also detect hbsab as low as 1iu/ml if i remember correct, so check with this too.

so after these tests the choices are:
hbv vaccine repeted monthly even for a year (they did this in italy to transplanted patients and 50% did hbsab), there are also cases like yours that had hbsab increase by hbv vaccine

try alinia+hbv vaccine

make biopsy with staining for hbsag/intraepatic hbvdna/cccdna so you will know 100% if there is any virus left in the liver or at least staining for hbsag because the others are made only at research centers

[Macola]

I just found out that our clinic for infectious diseases bought abbot architect last month. If it shows negative results on HBsAg, do you advice biopsy, or is it best to do the liver biopsy irrespective of the results by architect (because you said it will show for 100 %)?

[stef2011]

infecting my child in

hbv vaccine will stop any possible infection to others


regarding alinia: what if it turns out that there is no occult HBV?
you wasted some money because it has no sides or prevented flu/herpes if you happen to catch a flu/herpes virus
it is off label for hbv and not sold in europe except inside specialized hospital for hcv and stomach infections.not possible to get prescription in europe

consequences if there is no hbv at all and I use the drug?
already told you above, less money in ur pocket

there is no hbv at all?
only biopsy might tell 100%

, if it turns out to be an occult hbv
it is very difficult it is according to me but occult hbv is very dangerous, you don t see it clinically but it promotes liver cancer/cirrhosis despite all normal parameters and no hbv visible unless very low hbvdna.so it is very dangerous for HCC
all this damage is believed to be because of the hbsag mutants

[Macola]

I'll look into it, but first I have to find a good hepatologist in my country because I don't think I can buy the medicine without a proper prescription - to be honest, I heard about it from you for the first time. I have never used any medications regarding Hep b, the doctors said that since it was acute, I didn't need any, so all of this is my initiative because I AM REALLY afraid of infecting my child in case I have an occult HBV infection (if I decided to start a family). I have one more question regarding alinia: what if it turns out that there is no occult HBV? You said that it might make hbsab appear in case there is hbsab, but what are the possible consequences if there is no hbv at all and I use the drug? Am I too optimistic thinking that there is no hbv at all?

You have no idea how helpful you've been to me. Honestly, if it turns out to be an occult hbv - I can live with that; if it turns out that I am not a carrier - I would thank Good for the rest of my life. It is not knowing what exactly it is, that is driving me crazy.

[stef2011]

if you have very cheap ryanair flights in ur country the best and cheapest might be a filght to hamburg germany or ospedale maggiore milan or ospedale cisanello pisa.

before making additional tests you might even try 6 motnhs of nitazoxanide (alinia) and see if there are any changes.
it is active even if there are hbsag mutants theorically since it has a wide spectrum of activity on envelopped viruses so you wont make any harm and if it is wild type hbsag you will see hbsab appear

[Macola]

http://www.natap.org/2011/APSL/APSL_05.htm
I read this reviews on the method, and I pray for the most accurate results because BELIEVE ME there are no other methods other than elisa and eclia here - I checked all the labs  today. Maybe my other choice is to go abroad (probably Serbia or Bulgaria) to do the tests.
Here, They don't check our donated blood (or maybe occasionally), I know it for sure. Even if they do, the methods are old as the bible. Sometimes you have to wait for 4-5 months just to do the regular tests for hepatitis (I quote: "Sorry, come back in 4 to 5 months, we are out of reagents). Go privately if you don't want to wait". Never mind...
Thank you so much for everything

[stef2011]

you should find these machines where they test donated blood for transusions

as to the machines for hbvdna detection sensibility is usually 6-10iu/ml (1iu/ml about 5.8copies/ml)
abbott hbvdna realtime has sensibility of 10iu/ml (about 50-60copies/ml)

[stef2011]

this is an article from 2010
Researchers Identify Lab Test That Detects HBsAg with Mutations
Mutations in HBsAg can cloak or hide HBV infection in donated blood or in patients undergoing lab tests for hepatitis B. Researchers have been trying to develop more sensitive tests that can identify HBV in blood, even when HBsAg mutations are present.

Recently, a team of European researchers tested 147 blood samples containing HBsAg with varying mutations with two generations of a commercial HBsAg test (Enzygnost® HBsAg 5.0 and 6.0, Siemens Healthcare Diagnostics Products) to see which test was more effective.

The second test was more accurate in identifying HBsAg with mutations.

“The results showed that modifications in design of the assay (test) improved considerably the ability of the test to detect HBsAg mutants, and that difficulties in detecting such HBV variants should not be expected with the routine use of the test in diagnostic laboratories and in blood transfusion centers,” researchers wrote in the Journal of Medical Virology.

[stef2011]

dont know much about it but that machine runs many different type of tests so it also matters the kit used i guess, not just the machine itself
ask sensitivity of machine for occult hbv test hbsag and hbvdna to the most expert tech/doctor in the lab, they should know

[Macola]

I just found a lab where I can test my blood, and they use Roche Elecsys 2010. Is this sensitive enough, or is it the same as eclia and elisa?

[Macola]

Thank you very very very much for your help. By the way, I am from Macedonia.

[stef2011]

My HBsAg is expressed in iu/ml, and it is negative (less than 1.0 is negative).

probably yes, check if hbsag is made by architect or roches hbsag quant or siemens because only these can test for hbsag mutants
the test with less than 1.0 is negative is usually elisa or eclia, the hbsag tests about 20years old

the fact is that we are checking for possible occult hbv and maximum sensibility is needed.i is also possible to make a biopsy and check hbsag and hbvdna directly in liver cells, this of course is way much more sensible than a blood test, also cccdna would be very useful
hbsag can be positive in the liver and negative in the blood and the same can be for hbvnda

which country are you from?

[Macola]

OMG, but this is the only place in my country where this test can be done. People from the whole country come here to do the tests...
My HBsAg is expressed in iu/ml, and it is negative (less than 1.0 is negative). I suppose this is useless too?

[stef2011]

old useless tests have been used for you, copies/ml is not used anymore from many years results must be measure by iu/ml

when i was 18yo the test used had a sensitivity of 104copies/ml so more than 23 years ago the tests were better than the one used for you.
since you have to test for occult hepatitis they must detect very low quantity of hbvdna and the test you have used is totally useless on any hbv carrier, 400copies is way too much

could detect even one copy if it existed
bulls***, they can throw their test kit to the bin

you have to look for a test with sensitivity 6iu/ml, some tqman tests can reach this or make this test which is the most sensible in the world

http://www.ngi.com/
http://www.ngi.com/downloads/hbv_ultraqual.pdf