Colorimetric detection of norovirus by helicase-dependent amplification method based on specific primers integrated with HRPzyme

• Published in: Analytical and bioanalytical chemistry 2022-09, Vol.414 (23), p.6723-6733
• Main Authors: Lee, Jeong-Eun, Kim, Sol-A., Park, Hyun-Jin, Mun, Hyoyoung, Ha, Kwang-Soo, Shim, Won-Bo
• Format: Article
• Language: English
• Subjects: Amplification; Analysis; Analytical Chemistry; Biochemistry; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Colorimetry; DNA helicase; Food Science; Foodborne diseases; Gastroenteritis; Gene amplification; Illnesses; Laboratory Medicine; Methods; Monitoring/Environmental Analysis; Norovirus; Paper in Forefront; Transcription
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-022-04247-5

Noroviruses (NoVs) are the most common causes of epidemic gastroenteritis, responsible for at least 50% of all gastroenteritis outbreaks worldwide and significant causes of foodborne illness. In the USA, approximately 21 million illnesses attributable to NoVs have annually occurred. Therefore, there is a great demand to develop a rapid, low-cost, and accurate detection method for NoVs. This study first reported colorimetric helicase-dependent amplification (HDA) methods based on specific primers integrated with HRPzyme for the rapid and sensitive detection of NoV GI and GII. The colorimetric HDA methods exhibited a detection limit of 10 copies mL −1 of each NoV GI and GII and were confirmed to be specific to each NoV GI and GII. The period required to complete the HDA method was 2 h, including a step of RNA extraction and cDNA synthesis without expensive instruments such as a thermal cycler and detector. The cutoff value of the method for the oyster artificially inoculated with a known amount of NoV was all 10 2 copies g −1 for NoV GI and GII. Therefore, the HDA method developed in this study can be useful tool for the on-site detection of NoVs in food samples. Graphical abstract