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Original Article
Online Published: 08 Dec 2015
 


Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja.


Abstract
Designing a rapid, reliable and sensitive assay, for detection of hepatitis B virus and Hepatitis C virus variants by real-time PCR, is challenging at best. A recent approach for quantifying the viral load using the sensitive fluorescence principle, was used in this study. A total of 350 samples were collected from outpatient unit, CLRD. Complete Human HBV DNA and HCV sequences were obtained from the National Centre for Biotechnology Information (NCBI); primers and probes were designed and synthesized from core, surface and x region of Hepatitis B and UTR region of HCV. Real-time based detection was done, using standard kit and in-house generated standards and RT-PCR protocols. A standard curve was generated by using the Smart Cycler II software and serial dilution 102 to 108 of cloned viral regions, the calibration curve was linear in a range from 102 to108 cp/ml for both HBV and HCV, with R2 value of 0.999 and 0.995. Out of 100 predetermined HCV negative samples, 02 samples were found positive with in-house developed RT-PCR assay, the positivity of this sample was confirmed by sequencing the amplified product. Low cost of this assay procedure and précised sample volume will permit the assay to be implemented for routine screening of Hepatitis B and C virus mono-infection and co-infection using Real Time PCR , Nucleic acid Chip technology and Fluorescent End Point detection systems. 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 HBV and HCV virus monitor.

Key words: Hepatitis B virus, Hepatitis C virus, Nucleic acid test, Real-time PCR


 
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How to Cite this Article
Pubmed Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. J Med Allied Sci. 2011; 1(2): 69-73.

Web Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. https://jmas.in/?mno=210240 [Access: September 12, 2024].

AMA (American Medical Association) Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. J Med Allied Sci. 2011; 1(2): 69-73.


Vancouver/ICMJE Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. J Med Allied Sci. (2011), [cited September 12, 2024]; 1(2): 69-73.


Harvard Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja (2011) Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. J Med Allied Sci, 1 (2), 69-73.


Turabian Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. 2011. Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. Journal of Medical and Allied Sciences, 1 (2), 69-73.


Chicago Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. "Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry." Journal of Medical and Allied Sciences 1 (2011), 69-73.


MLA (The Modern Language Association) Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja. "Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry." Journal of Medical and Allied Sciences 1.2 (2011), 69-73. Print.


APA (American Psychological Association) Style

Y. Naresh Kumar, S. Rahmathulla, C. Madhavi, V. V. Ramachandra, S. Vishnupriya, Mohammed Aejaz Habeeb, M. N. Khaja (2011) Simultaneous detection of Hepatitis B virus and Hepatitis C virus in human plasma using Taq-man chemistry. Journal of Medical and Allied Sciences, 1 (2), 69-73.


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