QUANTITATIVE TEST FOR HBV BY RT-PCR

VIRUS DETECTED: Hepatitis B virus

INDICATIONS FOR ORDERING: Detection or presence of an acute or chronic HBV infection in a patient

METHOD USED: Real time quantitative polymerase chain reaction kits provided by Qiagen India for detection of HBV

SPECIMENS – Type, storage and transport

• Type: Non-diluted peripheral blood sample in an EDTA (lavender) tube. PLEASE AVOID HEPARINATED SAMPLES.

Whole Blood: 3 mL

• Transport: Specimens must be received within 48 hours of collection
• Storage: Serum Plasma: Separate specimens must be submitted when multiple tests are ordered.
• Unacceptable samples: Specimens collected in anticoagulants other than EDTA, less or no plasma serum in the EDTA vial

TEST DESCRIPTION: This is a viral DNA based real-time quantitative test, based on the amplification of specific regions of the pathogen genome. The viral genome is isolated from the peripheral blood serum and employed in the real-time PCR for amplification. In this assay, the amplified product is detected using the fluorescent dyes. The kit provides specific probes and primers that specifically to the amplified product. The fluorescence intensities are monitored which allow the detection and quantitation of the PCR product. The kit is provided with an internal control (IC) which is employed in the samples and positive and negative controls to check for possible PCR inhibition. The negative samples can be validated when the Ct value is >29. The PCR product is quantitated using the standard curve into IU/ml of samples:

The linear range is set between 3.8 – 4.0×10⁹ IU/ml. If HBV DNA detected in this range, the infection is reported. Below the sensitivity range, the infection may not be reported.

ANALYTICAL SENSITIVITY: The assay detects the presence of HBV at a value as low as 3.8 IU/ml (p = 0.05) calibrated against WHO 1^st International HBV DNA Standard. This means that there’s a 95% probability that 3.8 IU/ml will be detected, if present.

TURNAROUND TIME: 5 (five) days from the receipt of sample

QUALITY ASSURANCE & COMPETENCY TESTING: Regular interval quality checks and intra-lab comparisons are performed to ensure accurate and precise results. Highly trained personnel performing this test undergo regular competency assessments and attend regular conferences and educational programs in order to stay upbeat with the newer advances in the biology and diagnostics of the disease in question.

ACCREDIATION: NOT IN NABL SCOPE