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HEPATITIS B VIRUS VIRAL LOAD

Categoría: Infectious diseases.
Días hábiles de entrega: 10
 

EDTA blood, purple cap tube, serum, plasma.

Store and ship samples at temperatures between approximately 4-8 ° C; refrigerant gels or refrigerator ice can be used for it.

DNA extraction from the sample using commercial kits.

Real-time PCR using primers and SYBR® Green I No ROX Intercalators for the quantification of HBV DNA, which were designed from the sequence of the non-coding 5'NCR region of the hepatitis C virus, according to Carman, W. et al. 1990. For the quantification, we constructed a standard curve starting from the DNA extracted from the quantified serum of a positive HBV patient (positive control). DNA amplification is detected as an increase in fluorescence. A Step One thermal cycler from Applied Biosystems is used for the realization of the technique.

Positive control: sera from HBV DNA positive patients and with known viral load.

10 working days.

Negative: No amplification detected HBV DNA LOG: --

Positive: DNA is detected HBV XI / ml LOG: x HBV DNA

Limit of detection of the test: 100 IU / ml.

It is recommended that absolute values of viral load obtained by genomic amplification procedures be interpreted as any value within a range of 0.5 log (± 300 times) of the value obtained.

Genomik Laboratory has validated the test with positive and negative commercial controls; however, a negative result does not exclude the presence of HBV DNA below the sensitivity limit of the test, or the possible presence of PCR inhibitors. The reliability of the test is guaranteed by the use of systems for the prevention of contamination, in addition to the incorporation of an internal control of each test to avoid the appearance of false negatives.

Viral load tests measure the amount of HBV DNA (genetic material) circulating in the blood. A detectable viral load indicates that HBV is actively multiplying. When liver enzymes have an abnormal level, the higher viral load of HBV DNA appears to be associated with a greater severity of liver disease. Viral load tests are also helpful in indicating if treatment is being effective. People infected with hepatitis B should receive regular medical care. Biochemical liver tests should be repeated periodically (every 6-12 months). During treatment, the level of HBV DNA should also be monitored frequently to determine how the therapy is working. When taking interferon, side effects should be monitored by analyzing red blood cell counts, testing thyroid function, and evaluating the possible presence of depression. Carriers of inactive HBV should be tested for ALAT regularly. All HBV carriers should be tested for liver cancer. It is important to find a doctor who has knowledge about hepatitis B; hepatologists and gastroenterologists are specialized liver diseases.

Carman, W. et al. Vaccine-induced escape mutant of hepatitis B virus. Med Science, The Lancet 1990; 336: 352-329.



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