Application of two RT-PCR methods for detection of norovirus in market-sold oysters and norovirus genetic characteristic analysis, a survey conducted in Beijing.
10.3760/cma.j.cn112338-20210519-00411
- VernacularTitle:应用2种RT-PCR方法检测和分析北京市市场销售牡蛎中诺如病毒基因特征
- Author:
Han Qiu YAN
1
;
Yong Quan WANG
2
;
Hai Yang CUI
2
;
Bo JIN
2
;
Zhi Yong GAO
1
;
Quan Yi WANG
1
Author Information
1. Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control/Beijing Research Center for Preventive Medicine, Beijing 100013, China.
2. Xicheng District Center for Disease Control and Prevention, Beijing 100120, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Beijing;
Humans;
Norovirus/genetics*;
Ostreidae;
RNA, Viral/genetics*;
Real-Time Polymerase Chain Reaction;
Reproducibility of Results;
Reverse Transcriptase Polymerase Chain Reaction
- From:
Chinese Journal of Epidemiology
2022;43(1):92-97
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To evaluate the application of real-time RT-PCR and semi-nested RT-PCR in the detection of norovirus in oysters and analyzing the genetic characteristics of the isolates. Methods: Real-time fluorescent RT-PCR and semi-nested RT-PCR were used to detect norovirus GⅠ/GⅡ in fresh oysters collected from the markets in Beijing from November 2014 to October 2015. The detection rate of the parallel test was also analyzed. In addition, the reliability of semi-nested RT-PCR was evaluated by agreement rate and consistency test (Kappa value). The positive products of norovirus GⅠ/GⅡ capsid protein region gene by semi-nested RT-PCR were sequenced. Software BioEdit 7.0.9.0 was used for sequence alignment, and software Mega 6.0 was used to construct the evolutionary tree. Results: In 72 samples, the detection rate of norovirus was 31.94% (23/72) by real-time RT-PCR, 38.89% (28/72) by semi-nested RT-PCR and 48.61% (35/72) by parallel test. The coincidence rate of the two methods was 73.61%, a moderate degree (Kappa value =0.43). A total of 13 norovirus strains were successfully sequenced, and 11 strains (7 GⅡ.17 strains, 2 GⅡ. 4 Sydney_ 2012 strains, 1 GⅡ. 1 strain and 1 GⅡ. 21 strain) were obtained from norovirus positive samples by two RT-PCR methods, two strains (1 GⅡ. 17 strain and 1 GⅡ. 3 strain) were obtained from real-time RT-PCR negative samples which were positive for norovirus by semi-nested RT-PCR. The similarity between these strains and reference strains from diarrhea patients, environmental sewage, and shellfish products were 84.4% - 100.0%. Conclusions: The parallel test of norovirus in oysters by two RT-PCR methods can improve the detection rate and detect more genotypes. Norovirus strains in oysters were highly homologous with reference strains from diarrheal patients, environmental sewage, and shellfish products. Therefore, surveillance, prevention and control for norovirus should be carried out in people who have frequent contacts with oysters and related environments.
