Using Real-time PCR Assay to Quantify BKV and JCV

• Main Authors: Ying-Hsu Chen, 陳瀅旭
• Other Authors: Chun-Nan Lee
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/56539770011063253154

碩士 === 國立臺灣大學 === 醫學檢驗暨生物技術學研究所 === 97 === JC virus (JCV) and BK virus (BKV) are two species of Polyomaviridae that can infect humans. Primary infections usually occur during childhood and remain latent in the body. Infections from both viruses are generally asymptomatic in immunocompetent hosts. However, reactivation of these viruses may occur in immunosuppressed individuals and could cause severe diseases. This study intended to develop a rapid and highly sensitive method of diagnosis that could detect reactivation of JCV and BKV in immunosuppressed patients. In early stages, time-consuming and less sensitive methods, such as cell culture, electron microscopy, and immunofluorescence were used for the detection of human polyomavirus. Recently, polymerase chain reaction (PCR) and real-time PCR are widely used for detection of viral DNA in clinical diagnosis. The goal of this study is application of real-time PCR in detection and quantification of JCV and BKV. At first, we as established to monitor the efficiency of the assay and to avoid false negative results. The results showed that that the primers and probes of internal control did not interact with JCV and BKV. However, there were some interferences between the internal control and target genes. We analyzed plasma and urine samples of 79 bone marrow transplant patients by real-time PCR, 51 (64%) and 54 (68%) were detected as BKV positive, respectively. The results correlated well between plasma and urine samples. Then we analyzed the factors associated with hemorrhagic cystitis (HC) in bone marrow transplant patients. There were no relationship between HC and age, sex, diagnosis, or source of bone marrow transplant.