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

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 {"type":"entrez-nucleotide","attrs":{"text":"EU684022","term_id":"189176131"}}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.

Association of vitamin D receptor gene start codon [Fok 1] polymorphism with high myopia

High myopia caused primarily due to abnormal emmetropization and excessive axial ocular elongation is associated with sight-threatening ocular pathology. Muscular dysfunction of ocular ciliary muscles due to altered intracellular calcium levels can result in defective mechanotransduction of the eye and retinal defocus. The vitamin D3 receptor [VDR; a intracellular hormone receptor] is known to mediate calcium homestasis, influencing the development of myopia. In the present study, a total of 206 high myopia, 98 low myopia and 250 control samples were analyzed for VDR gene Fokl [exon 2 start codon] polymorphism using polymerase chain reaction-restriction fragment length polymorphism [PCR-FRLP] technique. High myopia patients revealed decrease in the frequency of ff homozygotes [8.3%] as compared to control group [14.0%], with a corresponding increase in frequency of FF homozygotes [68.9% in high myopia vs. 62.8% in controls]. The frequency of fallele carriers [Ff and ff] was increased in females of high myopia [35.6%] and low myopia cases [45.4%]. Elevated frequency of f allele was found only in early age at onset cases of high myopia [0.227] and later age at onset [10- 20 years] cases of low myopia [0.273] as well as in low myopia cases with parental consanguinity [0.458] [P 0.035; chi [2] = 6.692[*]]. The results suggest that VDR gene might not be playing a direct role in the development conferred by mechanical stress factors or growth/ development related factors through its role in calcium homestasis and regulation of ciliary muscle function