Improvement of nested reverse transcription-polymerase chain reaction (RT-PCR) with high specificity and sensitivity detection of sapovirus in food matrix

Soontag Jung; Daseul Yeo; Zhaoqi Wang; Seoyoung Woo; Yeeun Seo; Md Iqbal Hossain; Min Suk Rhee; Changsun Choi

doi:10.1016/j.jviromet.2022.114609

Improvement of nested reverse transcription-polymerase chain reaction (RT-PCR) with high specificity and sensitivity detection of sapovirus in food matrix

J Virol Methods. 2022 Nov:309:114609. doi: 10.1016/j.jviromet.2022.114609. Epub 2022 Aug 24.

Authors

Soontag Jung¹, Daseul Yeo¹, Zhaoqi Wang¹, Seoyoung Woo¹, Yeeun Seo¹, Md Iqbal Hossain¹, Min Suk Rhee², Changsun Choi³

Affiliations

¹ Department of Food and Nutrition, Chung-Ang University, 4726 Seodongdaero, Daedeok-myeon, Anseong-si, Gyeonggi-do 17546, Republic of Korea.
² Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea.
³ Department of Food and Nutrition, Chung-Ang University, 4726 Seodongdaero, Daedeok-myeon, Anseong-si, Gyeonggi-do 17546, Republic of Korea. Electronic address: cchoi@cau.ac.kr.

PMID: 36028003
DOI: 10.1016/j.jviromet.2022.114609

Abstract

Sapovirus (SaV) is a causative agent of human gastroenteritis in both community outbreaks and sporadic cases worldwide. Shellfish accumulate a variety of pathogens during filter feeding. In particular, the contamination of shellfish by SaV has caused several outbreaks. As reported previously, nested RT-PCR (nRT-PCR) has been widely used in clinical samples, but has not proven suitable for food samples, such as oysters. This study aimed to identify a primer set for the detection of SaV with high specificity and sensitivity in food samples. To accomplish this, primers were improved in RNA-dependent RNA polymerase (RdRp) regions of SaV whole genome sequences. The sensitivity of the improved nRT-PCR was 100-1000 times higher than that of previous nRT-PCR and > 10 times higher than that of the previous real-time RT-PCR assay. Notably, cross-reaction with other viruses or food matrices was not observed by the specificity test. This study improved the reliable primer set to detect SaV in various food matrices with high sensitivity.

Keywords: Food safety; Nested RT-PCR; Oyster; Sapovirus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Caliciviridae Infections*
Feces
Humans
RNA-Dependent RNA Polymerase
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Reverse Transcription
Sapovirus* / genetics
Sensitivity and Specificity

Substances

RNA-Dependent RNA Polymerase