Construction of Improved PCR Primer Set for the Detection of Human EntericAdenovirus41

Abstract

Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of HumanParvovirusB19

“…HuPaV-B19 검출용 conventional PCR primer 조합 6개를 대상으로 검출 민감도를 분석한 결과 한 번의 PCR에서는 1 ng/μL 이하에서 100 fg/μL 수준이었으며, nested PCR 결과 1 ng/μL에서 10 fg/μL 수준으로 나타났다( Table 2) (Schwarz et al, 1988;Mori et al, 1989;Brown et al, 1990;Gray et al, 1993;Bostic et al, 1999 (Kittigul et al, 2005;Cho, 2018…”