| Summary: | 碩士 === 中山醫學大學 === 生化暨生物科技研究所 === 95 === The genotyping of polyomaviruses JC virus (JCV) has been traditionally determined by restriction fragment length polymorphism analysis and by nucleotide sequencing. These genotypes are primarily associated with geographically based virus evolution. To investigate its implication in the pathogenesis and diagnosis of JCV infection, this study aims to further assess the genetic polymorphism in the regulatory region of VP1 of CY, TW-1, TW-2 and TW-3 genotypes which predominate in Taiwanese.
A 400-bp amplified fragment of the regulatory region of VP1 was detected in 20% of urine samples from healthy adults receiving no immunosuppressants (n = 50). Twenty-four out of sixty (40%) kidney transplantation patients on immunosuppression medication were found positive for the VP1 amplification of JCV DNA. The genotypes of these amplified DNA fragments were further analyzed by capillary electrophoresis. It was found that 80% (n = 10) of healthy adults carried one single genotype whereas the rest carried two genotypes. 21% and 66% of JCV-positive kidney transplantation patients (n =24) respectively carried one and two genotypes whereas the remaining showed multi-genotype. Our result is in sharp contrast to previous findings by restriction fragment length polymorphism analysis which only detected one single genotype. It is worth noting that the amount of DNA used by this approach is 0.5% less than that by PCR analysis, suggesting greater sensitivity of capillary electrophoresis analysis. Such a technique can thus provide a new insight into the clinical diagnosis of JCV infection. Moreover, it will facilitate our ability to develop an effective therapeutic strategy for complications associated with immunosuppressant-induced JCV mutation in kidney transplantation patients. It is conceivable that the technique can be further implemented in the study of drug-resistance mutation of viruses such as BKV, HBV, HCV, HIV, etc.
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