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Human Sapovirus (HuSaV) is one of the major causes of acute gastroenteritis in humans, and it is used as a molecular diagnostic technique based on polymerase chain reaction (PCR) from humans, food, shellfish, and aquatic environments. In this study, the HuSaV diagnosis technique was used in an aquatic environment where a number of PCR inhibitors are included and pathogens, such as viruses, are estimated to exist at low concentration levels. HuSaV-specific primers are improved to detect 38 strains registered in the National Center for Biotechnology Information (NCBI). The established optimal condition and the composition, including the RT-nested PCR primers and SL<SUP>®</SUP> Non-specific reaction inhibitor, were found to have 100 times higher sensitivity based on HuSaV plasmid than the previously reported methods (100 ag based on HuSaV plasmid 1 ng/μL). Through an artificial infection test, the developed method was able to detect at least 1 fg/μL of HuSaV plasmid contaminated with total nucleic acid extracted from groundwater. In addition, RT-nested PCR primer sets for HuSaV detection can react, and a positive control is developed to verify false positives. This study is expected to be used as a HuSaV monitoring method in the future and applied to the safety response to HuSaV from water environments.
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참고문헌 (27)
참고문헌 신청
Botes M / 2013 / Application of quantitative PCR for the detection of microorganisms in water / Anal Bioanal Chem 405:91~108
Bhullar SS / 2014 / Determination of viral load by quantitative real-time PCR in herpes simplex encephalitis patients / Intervirology 57:1~7
조규봉 / 2018 / Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of Human Parvovirus B19 / 대한의생명과학회지 24(4):390~397
조규봉 / 2018 / Construction of Improved PCR Primer Set for the Detection of Human Enteric Adenovirus 41 / 대한의생명과학회지 24(3):230~238
Cho SR / 2018 / Pathogen surveillance of acute viral gastroenteritis in Korea in 2017 / Public Health Weekly Report 11:1374~1380
Bhullar SS / 2014 / Determination of viral load by quantitative real-time PCR in herpes simplex encephalitis patients / Intervirology 57:1~7
조규봉 / 2018 / Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of Human Parvovirus B19 / 대한의생명과학회지 24(4):390~397
조규봉 / 2018 / Construction of Improved PCR Primer Set for the Detection of Human Enteric Adenovirus 41 / 대한의생명과학회지 24(3):230~238
Cho SR / 2018 / Pathogen surveillance of acute viral gastroenteritis in Korea in 2017 / Public Health Weekly Report 11:1374~1380
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