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Use of biochemical miniaturized galleries, rRNA based lateral flow assay and Real Time PCR for Cronobacter spp. confirmation.
- Source :
-
Food microbiology [Food Microbiol] 2018 Dec; Vol. 76, pp. 189-195. Date of Electronic Publication: 2018 May 21. - Publication Year :
- 2018
-
Abstract
- Identification of Cronobacter represent a major challenge for laboratories testing powdered infant formula (PIF). In the present study, two biochemical galleries and three molecular methods have been applied to confirm 276 Cronobacter spp. and non-Cronobacter isolates from different sources. Using the latest database of API 20 E and ID 32 E biochemical miniaturized kits, 53% and 78% of the isolates were identified respectively. From the available results, total accuracy for Cronobacter detection was in 97% (API 20 E) and 99% (ID 32 E). The three molecular methods were based on rRNA based lateral flow, Real Time PCR combined with either a hybridization or hydrolysis probe. For all three methods total accuracy was more than 99%. A pilot trial using Next Generation Sequencing (NGS) correctly identified 58 out of 66 isolates (88%) in DNA mixtures. The results indicate that the commercially available approaches such as ID 32 E, rRNA based lateral flow and Real Time PCR are all suitable for Cronobacter identification at the genus level. The NGS method may become a suitable alternative in the future, provided that the sequence database is improved.<br /> (Copyright © 2018 Elsevier Ltd. All rights reserved.)
- Subjects :
- Bacterial Typing Techniques
Biochemistry instrumentation
Cronobacter classification
Cronobacter isolation & purification
Data Accuracy
Food Contamination analysis
Food Microbiology methods
Humans
Infant
Infant, Newborn
Miniaturization
Paper
Phylogeny
Reagent Kits, Diagnostic
Reagent Strips analysis
Biochemistry methods
Cronobacter genetics
DNA, Bacterial genetics
Infant Formula microbiology
RNA, Ribosomal, 16S genetics
Real-Time Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9998
- Volume :
- 76
- Database :
- MEDLINE
- Journal :
- Food microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 30166140
- Full Text :
- https://doi.org/10.1016/j.fm.2018.05.008