Multi-enzyme isothermal rapid amplification assay for the detection of Escherichia coli O157∶H7
10.19428/j.cnki.sjpm.2022.22048
- VernacularTitle:多酶恒温核酸快速扩增法检测大肠杆菌O157∶H7
- Author:
Shujuan WANG
1
;
Yiling FAN
1
;
Zhen FENG
1
;
Bo JIANG
1
;
Minghui SONG
1
;
Qiongqiong LI
1
;
Hao LIU
1
;
Feng QIN
1
;
Meicheng YANG
1
Author Information
1. NMPA Key Laboratory for Testing Technology of Pharmaceutical Microbiology，Shanghai Institute for Food and Drug Control， Shanghai 201203， China
- Publication Type:Journal Article
- Keywords:
multienzyme isothermal rapid amplification;
metal-organic framework immunomagnetic bead;
rfbE gene;
Escherichia coli O157∶H7
- From:
Shanghai Journal of Preventive Medicine
2022;34(6):511-518
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveA rapid enrichment and detection method for Escherichia coli O157∶H7 was developed by using multienzyme isothermal rapid amplification （MIRA） fluorescence method combined with metal organic frameworks immunomagnetic beads. MethodsUsing rfbE gene as the target， the primers， probes and reaction system were screened， and the specificity， sensitivity and practical application of this method were investigated. ResultsThe detection limit of Escherichia coli O157∶H7 was 1.18×105 CFU‧mL-1， and the detection limit of DNA concentration was 9 pg‧μL-1. The detection process was completed in 20 minutes. The test results of 47 strains （24 target strains and 23 non-target strains） were consistent with real-time PCR （RT-PCR）. ConclusionA method based on metal-organic framework immunomagnetic beads enrichment combined with MIRA assay is developed in this study. The method is simple， rapid and suitable for rapid enrichment and detection of Escherichia coli O157∶H7 in food.
