Development of an isothermal amplification-based assay for the rapid detection of Cronobacter spp.

被引：15

作者：

Liu, Siying ^{[1
]}

Geng, Yunyun ^{[1
,2
]}

Liu, Libing ^{[3
,4
]}

Sun, Xiaoxia ^{[3
,4
]}

Shao, Jingyu ^{[1
]}

Han, Beibei ^{[1
]}

Wang, Jianchang ^{[3
,4
]}

Tan, Ke ^{[1
]}

机构：

[1] Hebei Normal Univ, Coll Life Sci, Key Lab Anim Physiol Biochem & Mol Biol Hebei Pro, Shijiazhuang 050024, Hebei, Peoples R China

[2] Hebei Univ Chinese Med, Coll Basic Med, Shijiazhuang 050020, Hebei, Peoples R China

[3] Hebei Entry Exit Inspect & Quarantine Bur, Ctr Inspect & Quarantine, Shijiazhuang 050051, Hebei, Peoples R China

[4] Hebei Acad Inspect & Quarantine Sci & Technol, Shijiazhuang 050051, Hebei, Peoples R China

来源：

JOURNAL OF DAIRY SCIENCE | 2018年 / 101卷 / 06期

基金：

中国博士后科学基金;

关键词：

Cronobacter spp; recombinase polymerase amplification; real-time recombinase polymerase amplification; real-time PCR; ENTEROBACTER-SAKAZAKII; PROPIDIUM MONOAZIDE; MILK; MENINGITIS; PATHOGEN; PROBE; GENE;

D O I：

10.3168/jds.2017-13931

中图分类号：

S8 [畜牧、动物医学、狩猎、蚕、蜂];

学科分类号：

0905 ;

摘要：

Cronobacter spp. is an opportunistic pathogen that is associated with rare but life-threatening neonatal infections resulting from the consumption of contaminated powdered infant formula milk (PIF). In the present study, we developed recombinase polymerase amplification (RPA) and real-time RPA for the detection of Cronobacter spp. in PIF for the first time by targeting the ompA gene. The specificity and sensitivity of the RPA and real-time RPA were validated and the practical applicability of these methods for the detection of Cronobacter spp. in artificially contaminated PIF samples was proved by comparing their reaction time, sensitivity, and efficacy with those of real-time PCR and the Chinese traditional method. The RPA and real-time RPA assays reduced the analysis time to less than 15 min and the results were as reliable as those of real-time PCR. Taken together, the RPA and realtime RPA assays served as fast, reliable, and sensitive techniques for the detection of Cronobacter spp.

引用

页码：4914 / 4922

页数：9

共 25 条

[1] Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus [J].

Abd El Wahed, Ahmed ;

Weidmann, Manfred ;

Hufert, Frank T. .

JOURNAL OF CLINICAL VIROLOGY, 2015, 69 :16-21

[2] Recombinase Polymerase Amplification Assay for Rapid Diagnostics of Dengue Infection [J].

Abd El Wahed, Ahmed ;

Patel, Pranav ;

Faye, Oumar ;

Thaloengsok, Sasikanya ;

Heidenreich, Doris ;

Matangkasombut, Ponpan ;

Manopwisedjaroen, Khajohnpong ;

Sakuntabhai, Anavaj ;

Sall, Amadou A. ;

Hufert, Frank T. ;

Weidmann, Manfred .

PLOS ONE, 2015, 10 (06)

[3] Rapid Detection and Simultaneous Genotyping of Cronobacter spp. (formerly Enterobacter sakazakii) in Powdered Infant Formula Using Real-time PCR and High Resolution Melting (HRM) Analysis [J].

Cai, Xian-Quan ;

Yu, Hai-Qiong ;

Ruan, Zhou-Xi ;

Yang, Lei-Liang ;

Bai, Jian-Shan ;

Qiu, De-Yi ;

Jian, Zhi-Hua ;

Xiao, Yi-Qian ;

Yang, Jie-Yang ;

Thanh Hoa Le ;

Zhu, Xing-Quan .

PLOS ONE, 2013, 8 (06)

[4]

CFDA, 2016, GB4789402016 CFDA

[5] Development and application of a sensitive, rapid, and reliable immunomagnetic separation-PCR detection method for Cronobacter spp. [J].

Chen, Qiming ;

Li, Yuanhong ;

Tao, Tingting ;

Bie, Xiaomei ;

Lu, Fengxia ;

Lu, Zhaoxin .

JOURNAL OF DAIRY SCIENCE, 2017, 100 (02) :961-969

[6] Mining for sensitive and reliable species-specific primers for PCR for detection of Cronobacter sakazakii by a bioinformatics approach [J].

Chen Qiming ;

Tao Tingting ;

Bie Xiaomei ;

Lu Yingjian ;

Lu Fengxia ;

Zhai Ligong ;

Lu Zhaoxin .

JOURNAL OF DAIRY SCIENCE, 2015, 98 (08) :5091-5101

[7]

Drudy D, 2006, CLIN INFECT DIS, V42, P996, DOI 10.1086/501019

[8]

FDA, 2002, IS EN ENT SAK DEH IN

[9] Rapid detection of Cronobacter sakazakii by real-time PCR based on the cgcA gene and TaqMan probe with internal amplification control [J].

Hu, Shuangfang ;

Yu, Yigang ;

Li, Rong ;

Wu, Xinwei ;

Xiao, Xinglong ;

Wu, Hui .

CANADIAN JOURNAL OF MICROBIOLOGY, 2016, 62 (03) :191-200

[10] Cronobacter: an emerging opportunistic pathogen associated with neonatal meningitis, sepsis and necrotizing enterocolitis [J].

Hunter, C. J. ;

Bean, J. F. .

JOURNAL OF PERINATOLOGY, 2013, 33 (08) :581-585

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