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Validation of a PCR method for the detection Campylobacter spp. in milk powder

Nguyen Do Phuc Nguyen Ly Hoang Ngan Le Thi Hien Ngo Thanh Phong Tran Bao Tram Nguyen Ngoc Thanh Hien Nguyen Thi Thanh Truc Hoang Hoai Phuong
Published 09/21/2022

Article Details

How to Cite
Nguyen Do Phuc, Nguyen Ly Hoang Ngan, Le Thi Hien, Ngo Thanh Phong, Tran Bao Tram, Nguyen Ngoc Thanh Hien, Nguyen Thi Thanh Truc, Hoang Hoai Phuong. "Validation of a PCR method for the detection Campylobacter spp. in milk powder". Vietnam Journal of Food Control. vol. 5, no. 3, pp. 342-348, 2022
PP
342-348
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314

Main Article Content

Abstract

An alternative PCR method for rapid detection of Campylobacter spp. in milk powder was validated according to TCVN 12365-2:2018 (ISO 16140-2:2016). In this study, performed with paired study, validation examines the following parameters: (1) sensitivity (sensitivity for the alternative method: SEalt, sensitivity for the reference method: SEref, relative trueness: RT, false positive ratio for the alternative method: FPR), (2) relative level of detection (RLOD) and (3) inclusivity and exclusivity. The result showed that: sensitivity for the alternative method: 85.7%, sensitivity for the reference method 92.8%, relative trueness: 85%, false positive ratio for the alternative method: 16.7%, relative level of detection: 0,859 CFU/25g. For inclusitivity testing, two reference strains were detected. For exclusitivity testing, all of 20 strains were not detected. All the parameters tested were met according to TCVN 12365-2:2018 (ISO 16140-2: 2016) requirements.

Keywords:

Campylobacter spp., validation, PCR, milk powder, ISO 16140-2:2016

References

[1]. TCVN 12365-2:2018 (ISO 16140-2:2016): Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method.
[2]. ARL 04, Supplementary requirement for accreditation in the field of biological testing, Accreditation Office, 2 January 2020.
[3]. Hui Y. H., Pierson D., Richard Gorham J., Foobborne Disaese Hand Book, Second Sdition, volume 1, pp.83-106, 2001. Microbiology of the food chain - Horizontal method for detection and enumeration of Campylobacter spp. - Part 1: Detection method
[4]. ISO 10272-1: 2017: Microbiology of the food chain - Horizontal method for detection and enumeration of Campylobacter spp. - Part 1: Detection method
[5]. Protocol for PCR ampliction of Campylobacter jejuni and C. coli recommended by the EURL-AR 2nd version - November 2013.

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