Establishment And Application Of High - Throughput Detection Method For Human Polyoma Virus Based On MassARRAY

Human polyomaviruses (HPyVs) are widely distributed among the population worldwide. In the early years, BKPyV and JCKPyV are the only known members of the Polyimaviridae family of viruses of human origin, which can cause Polyomavirus-induced nephropathy and progressive multifocal leukoencephalopathy in human, respectively, in the context of severe immunosuppression and other predisposing factors. Over the past seven years,10novel HPyVs has been discovered, and the pathogenic roles of them still remain unclear, with the exception of MCPyV which causes Merkel Cell Carcinoma and TSPyV which leads to Trichodysplasia spinulosa. HPyVs were frequently detected in human stools or gastrointestinal tract organs, additionally, the three newest members of HPyVs-MWPyV, STLPyV and HPyV12-were all initially identified from fecal or gastrointestinal specimens, which brings attention to the relationship between HPyVs and human gastrointestinal illness, especially diarrhea.By using the matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry platform known as MassARRAY (Sequenmon Inc., USA), we designed and optimized a high-throughput detection method based on multiple PCR which can diagnose all12HPyVs simultaneously. We conducted a case-control study utilizes this method, and examined419stool samples from211children with diarrhea and208asymptomatic control subjects. Five HPyV species were detected in the fecal samples, including KIPyV, WUPyV, MCPyV, MWPyV and STLPyV. HPyV was not more prevalent among the case group than the control group; the difference in viral loads of MWPyV between the two groups was not statistically significant (p>0.05); our data did not support a causative role of HPyV in gastroenteritis. We identified16whole HPyV genome sequences and are the first to report the whole genome sequences of KIPyV, MCPyV, MWPyV and STLPyV isolated in China. This study provides the basis for in-depth etiological research of HPyVs and evolutional researches of their genomes.