Development of a novel and rapid polymerase spiral reaction (PSR) assay to detect Salmonella in pork and pork products

被引:29
作者
Momin, Kasanchi M. [1 ]
Milton, Arockiasamy Arun Prince [1 ]
Ghatak, Sandeep [1 ]
Thomas, Shiny C. [2 ]
Priya, Govindarajan Bhuvana [1 ]
Das, Samir [1 ]
Shakuntala, Ingudam [1 ]
Sanjukta, Rajkumari [1 ]
Puro, Kekungu-u [1 ]
Sen, Arnab [1 ]
机构
[1] Indian Council Agr Res ICAR Res Complex NEH Reg, Div Anim Hlth, Res Complex NEH Reg, Umiam 793103, Meghalaya, India
[2] Assam Don Bosco Univ, Sch Life Sci, Gauhati, Assam, India
关键词
Salmonella; Polymerase spiral reaction; Pork; Pathogen detection; MEDIATED ISOTHERMAL AMPLIFICATION; CHICKEN MEAT; PCR ASSAY; LAMP; CULTURE; PREVALENCE; ENRICHMENT; SAMPLES; SPP; FOOD;
D O I
10.1016/j.mcp.2020.101510
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The polymerase spiral reaction (PSR), a novel isothermal method for targeted DNA amplification, was effectively applied to detect Salmonella in artificially spiked pork. The specificity of the developed PSR was tested using 16 Salmonella and 15 non-Salmonella strains. The PSR assay was 10-fold more sensitive than conventional end-point PCR, having a sensitivity comparable to real-time PCR. The limit of detection of the developed assay was 4 x 10(3) per gram of pork without enrichment and 4 CFU per gram after a 6 h enrichment. The detection of 4 CFU per gram of pork was achieved within 8 h. The PSR assay was successful, and accurate in comparison to microbiological methods, in detecting Salmonella in 11 of 76 commercial pork samples. Therefore the positive predictive value, negative predictive value and accuracy rate of the developed assay were 100%. Considering its rapidity, user-friendliness, simplicity, cost-effectiveness and equipment-free nature, this PSR assay is a promising tool for the food industry for the detection of Salmonella and prevention of Salmonella outbreaks and recalls.
引用
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页数:9
相关论文
共 38 条
[1]  
[Anonymous], SALMONELLAE FOODS FE
[2]  
[Anonymous], 2002, Microbiology of food and animal feeding stuffs. Horizontal method for the detection of Salmonella spp
[3]   Development and evaluation of real-time loop mediated isothermal amplification assay for rapid and sensitive detection of Salmonella spp. in chicken meat products [J].
Arunrut, Narong ;
Kiatpathomchai, Wansika ;
Ananchaipattana, Chiraporn .
JOURNAL OF FOOD SAFETY, 2018, 38 (06)
[4]   Sensitive and specific detection of Cryptosporidium species in PCR-negative samples by loop-mediated isothermal DNA amplification and confirmation of generated LAMP products by sequencing [J].
Bakheit, Mohammed A. ;
Torra, Dena ;
Palomino, Lily A. ;
Thekisoe, Oriel M. M. ;
Mbati, Peter A. ;
Ongerth, Jerry ;
Karanis, Panagiotis .
VETERINARY PARASITOLOGY, 2008, 158 (1-2) :11-22
[5]   Virulence genes detection of Salmonella serovars isolated from pork and slaughterhouse environment in Ahmedabad, Gujarat [J].
Chaudhary, J. H. ;
Nayak, J. B. ;
Brahmbhatt, M. N. ;
Makwana, P. P. .
VETERINARY WORLD, 2015, 8 (01) :121-124
[6]   Global Trends in Typhoid and Paratyphoid Fever [J].
Crump, John A. ;
Mintz, Eric D. .
CLINICAL INFECTIOUS DISEASES, 2010, 50 (02) :241-246
[7]   Rapid visual isothermal nucleic acid-based detection assay of Brucella species by polymerase spiral reaction [J].
Das, A. ;
Kumar, B. ;
Chakravarti, S. ;
Prakash, C. ;
Singh, R. P. ;
Gupta, V. ;
Singh, K. P. ;
Agrawal, R. K. ;
Chaturvedi, V. K. ;
Abhishek ;
Shrinet, G. .
JOURNAL OF APPLIED MICROBIOLOGY, 2018, 125 (03) :646-654
[8]   Rapid detection of Pseudomonas aeruginosa targeting the toxA gene in intensive care unit patients from Beijing, China [J].
Dong, Derong ;
Zou, Dayang ;
Liu, Hui ;
Yang, Zhan ;
Huang, Simo ;
Liu, Ningwei ;
He, Xiaoming ;
Liu, Wei ;
Huang, Liuyu .
FRONTIERS IN MICROBIOLOGY, 2015, 6
[9]   Polymerase spiral reaction (PSR): a novel, visual isothermal amplification method for detection of canine parvovirus 2 genomic DNA [J].
Gupta, Vikas ;
Chakravarti, Soumendu ;
Chander, Vishal ;
Majumder, Saurabh ;
Bhat, Shabir Ahmad ;
Gupta, Vivek Kumar ;
Nandi, Sukdeb .
ARCHIVES OF VIROLOGY, 2017, 162 (07) :1995-2001
[10]   Loop-mediated isothermal amplification for the rapid detection of Salmonella [J].
Hara-Kudo, Y ;
Yoshino, M ;
Kojima, T ;
Ikedo, M .
FEMS MICROBIOLOGY LETTERS, 2005, 253 (01) :155-161