Real Time-PCR HBV-DNA Analysis: Significance and First Experience in Armed Forces

Chopra, GS; Gupta, PK; Anand, Anil C.; Varma, Prerna; Nair, Velu; Rai, Ramji

doi:10.1016/s0377-1237(05)80161-3

Cited by 8 publications

(9 citation statements)

References 15 publications

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“…The sensitivity of the TaqMan procedure for quantifying viral DNA has already been proved by various studies;[23161819] however, they are being limited by certain aspects such as limited samples, designing of primer, and probes set from regions of unconserved nature.…”

Section: Discussionmentioning

confidence: 99%

“…We also made the entire quantitative procedure in a low cost and rapid manner, which will be helpful in detecting HBV in patient′s blood in less time and accurate manner. The sensitivity of the TaqMan procedure for quantifying viral DNA has already been proved by various studies;[ 2 3 16 18 19 ] however, they are being limited by certain aspects such as limited samples, designing of primer, and probes set from regions of unconserved nature.…”

Section: Discussionmentioning

confidence: 99%

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A latest and promising approach for prediction of viral load in hepatitis B virus infected patients

Naresh¹,

Syed²,

Chandra³

et al. 2011

Indian J Hum Genet

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INTRODUCTION:Designing a rapid, reliable and sensitive assay for detection of hepatitis B virus (HBV) variants by real-time PCR is challenging at best. A recent approach for quantifying the viral load using a sensitive fluorescent principle was brushed in this study.MATERIALS AND METHODS: A total of 250 samples were collected from the outpatient unit, CLRD. Complete Human HBVDNA sequences (n = 944) were selected from the National Centre for Biotechnology Information (NCBI), primers and probes were designed and synthesized from the core, surface, and x region. Real-time based quantification was carried out using a standard kit and in-house generated standards and RT-PCR protocols.RESULTS AND DISCUSSION:The standard calibration curve was generated by using serial dilution 102 to 108. The calibration curve was linear in a range from 102 to 108 copies/ml, with an R2 value of 0.999. Reproducibility as measured by dual testing of triplicates of serum samples was acceptable, with coefficients of variation at 6.5%, 7.5%, and 10.5%. Our results showed that amplification performance was good in the case of the x-region-based design (98%). Out of 100 negative samples screened by enzyme linked immunosorbent assay and the standard RT-PCR kit, one sample was detected as positive with the in-house developed RT-PCR assay, the positivity of the sample was confirmed by sequencing the amplified product, NCBI accession EU684022.CONCLUSION:This assay is reproducible showing limited inter- and intra-assay variability. We demonstrate that the results of our assay correlated well with the standard kit for the HBV viral load monitor.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A latest and promising approach for prediction of viral load in hepatitis B virus infected patients

Naresh¹,

Syed²,

Chandra³

et al. 2011

Indian J Hum Genet

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“…These all samples were negative for HBV-DNA PCR. Negative result for HBV/DNA on PCR indicates that these patients have no titer of viral DNA in their serum [12].…”

Section: Confirmation Of Hbv-dna Through Pcrmentioning

confidence: 99%

Investigation of an inflammatory viral disease HBV in cardiac patients through polymerase chain reaction

Saher¹,

Rehman²,

Qureshi³

et al. 2012

ABB

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In the present study 100 cardiac patients were randomly selected from the cardiology ward, Allied Hospital Faisalabad, Pakistan. All the selected cases were analysed for different parameters like Hepatitis B surface Antigen (HbsAg), Bilirubin, Alkaline phosphatase, serum glutamic pyruvic transaminase, and serum glutamic-oxaloacetic transaminase. Out of total 16% patients were lying in the age of 21-30 year, 25% in the age of 31-40 year, 35% in the age of 41-50 year, 19% in the age of 51-60 years and 5% patients in the age of 61-70 years. No subject was found positive in age group 21-30 years patients. 35% patients have higher value of SGPT while, other 26% were with higher value of SGOT. Rest of the 32 and 24% have higher ALP and Bilirubin levels, respectively. Assay profile revealed that ALP level was increased with increasing age, body mass index, Creactive protein, diabetes, smoking, sex, serum uric acid, lead, cadmium, hypercholesterolemia, lesion of liver and cardiovascular disease. The serum of the eight HbsAg positive cases were tested for the presence of HBV through PCR and no sample was found positive. At the end of the study, PCR amplified samples were run on 1.5% agarose gel to confirm the case

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“…Amplification reactions were performed in 30 µl HBV RG master mix (tampon, dNTP, primer, probe and enzyme) and 50 µl reaction mixtures containing 20 µl templates DNA. The amplification protocol consisted of an initial denaturation at 95°C for 10 min, followed by 45 cycles of denaturation at 95°C for 15 seconds, annealing at 55°C for 30 seconds, and extension at 72°C for 15 seconds in a Rotor Gene 2000 device [12]. The copy limit of the test was 26 copies/ml.…”

Section: Hbv Dna Determination By Real Time Pcrmentioning

confidence: 99%

Correlation of Hepatitis B Virus Surface Antigen Titer with Hepatitis Delta Virus Replication Level and Histological Activity in Chronic Hepatitis D

Şimşek¹

2018

BJSTR

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We have analyzed the relationships between quantitative HDVRNA, HBsAg and HBV DNA concentrations and biochemical activity, histological activity index and fibrosis stage in 46 patients with chronic hepatitis D (CHD) and 87 with chronic hepatitis B (CHB), including 33 positive for HBeAg.Methods: All patients were naïve to treatment. HBsAg concentrations were measured using Abbott Architect kits and HDVRNA by real time PCR. Results and Conclusions:HBsAg and HDVRNA concentrations were significantly and positively correlated (p=0.043). HDV viremia varied greatly, with HDVRNA titer not correlated with biochemical or histological activity or fibrosis stage. Serum transaminase concentrations and histological findings did not differ between HBVDNA-negative and -positive patients with CHD (p>0.05), and HBsAg concentrations did not differ significantly in HBVDNA-positive and -negative patients (p>0.05). HBsAg levels were significantly higher in HBeAg-positive patients with CHD than in those with HBeAg-negative CHB but were similar in patients with CHD and HBeAg-negative CHB. In CHB patients, there was a positive correlation between serum HBVDNA and HBsAg concentrations (p=0.002). As a result, histological activity and fibrosis stage were significantly higher in CHD than CHB patients, but were not influenced by the levels of HDV and HBV viremia in CHD. HBsAg titers were highest in patients with HBeAg-positive CHB, followed by patients with CHD, and lowest in HBeAg-negative CHB patients.

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Real Time-PCR HBV-DNA Analysis: Significance and First Experience in Armed Forces

Cited by 8 publications

References 15 publications

A latest and promising approach for prediction of viral load in hepatitis B virus infected patients

A latest and promising approach for prediction of viral load in hepatitis B virus infected patients

Investigation of an inflammatory viral disease HBV in cardiac patients through polymerase chain reaction

Correlation of Hepatitis B Virus Surface Antigen Titer with Hepatitis Delta Virus Replication Level and Histological Activity in Chronic Hepatitis D

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