Application of in situ loop-mediated isothermal amplification method for detection of Salmonella in foods

Abstract In situ loop-mediated isothermal amplification ( in situ LAMP) as a novel technique for detection of food-borne pathogens, has been successfully applied to detect Salmonella in artificially contaminated eggshells. Escherichia coli C600 was used as a negative control in specificity experiment. The sensitivity of the in situ LAMP assay was confirmed by the test in serial 10-fold dilutions of Salmonella cells, and the detection limit of this method may reach 1 CFU/cm 2 on eggshells. Compared with traditional culture methods and PCR-based methods, the in situ LAMP assay is advantageous on rapidity, high specificity, less time consumption and ease in operation. This is the first study carried out to apply the in situ LAMP method in practical food-borne pathogens detection. The results indicate that this method has great potential for use in food safety laboratories.

[1]  Reiner Helmuth,et al.  A real-time PCR for the detection of Salmonella Enteritidis in poultry meat and consumption eggs. , 2007, Journal of microbiological methods.

[2]  R. Weigel,et al.  Comparison of API 20E and invA PCR for Identification of Salmonella enterica Isolates from Swine Production Units , 2006, Journal of Clinical Microbiology.

[3]  C. Surette,et al.  Quantification of Salmonella spp. in composted biosolids using a TaqMan qPCR assay. , 2007, Journal of microbiological methods.

[4]  W. Holben,et al.  Quantification of mRNA in Salmonella sp. seeded soil and chicken manure using magnetic capture hybridization RT-PCR. , 2007, Journal of microbiological methods.

[5]  E. Isogai,et al.  Detection of Salmonella invA by isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) in Zambia. , 2005, Comparative immunology, microbiology and infectious diseases.

[6]  C. Yamashiro,et al.  The evaluation of a fluorogenic polymerase chain reaction assay for the detection of Salmonella species in food commodities. , 1997, International journal of food microbiology.

[7]  A. Thaler,et al.  Food Safety and Inspection Service Policy for Control of Poultry Contaminated by Digestive Tract Contents: A Review , 2005 .

[8]  N. Nakasone,et al.  Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method. , 2005, FEMS microbiology letters.

[9]  S A McEwen,et al.  Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. , 1992, Molecular and cellular probes.

[10]  C Tirado,et al.  WHO surveillance programme for control of foodborne infections and intoxications: preliminary results and trends across greater Europe. World Health Organization. , 2001, The Journal of infection.

[11]  R. Savan,et al.  Sensitive and Rapid Detection of Edwardsiellosis in Fish by a Loop-Mediated Isothermal Amplification Method , 2004, Applied and Environmental Microbiology.

[12]  Nobuyasu Yamaguchi,et al.  Detection of Bacteria Carrying the stx2 Gene by In Situ Loop-Mediated Isothermal Amplification , 2003, Applied and Environmental Microbiology.

[13]  R. Xiang,et al.  A loop-mediated isothermal amplification method targets the phoP gene for the detection of Salmonella in food samples. , 2009, International journal of food microbiology.

[14]  I. Biunno,et al.  Technical note: Improved method for rapid DNA extraction of mastitis pathogens directly from milk. , 2006, Journal of dairy science.

[15]  R. Shashidhar,et al.  Rapid, sensitive, and validated method for detection of Salmonella in food by an enrichment broth culture - nested PCR combination assay. , 2008, Molecular and cellular probes.

[16]  Y. Hara-Kudo,et al.  Detection of Salmonella enterica in Naturally Contaminated Liquid Eggs by Loop-Mediated Isothermal Amplification, and Characterization of Salmonella Isolates , 2005, Applied and Environmental Microbiology.

[17]  G. Dougan,et al.  Evaluation of Salmonella typhimuriumMutants in a Model of Experimental Gastroenteritis , 1999, Infection and Immunity.

[18]  T. Notomi,et al.  Loop-mediated isothermal amplification of DNA. , 2000, Nucleic acids research.

[19]  Lin Li,et al.  Specific and rapid detection of foodborne Salmonella by loop-mediated isothermal amplification method , 2008 .

[20]  Takashi Kawana,et al.  Tolerance of loop-mediated isothermal amplification to a culture medium and biological substances. , 2007, Journal of biochemical and biophysical methods.

[21]  C. Braden,et al.  Salmonella enterica serotype Enteritidis and eggs: a national epidemic in the United States. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[22]  T. Notomi,et al.  Accelerated reaction by loop-mediated isothermal amplification using loop primers. , 2002, Molecular and cellular probes.

[23]  Kaiser Jamil,et al.  Rapid detection of food-borne pathogens by using molecular techniques. , 2005, Journal of medical microbiology.

[24]  Lin Li,et al.  Quinolone-resistance in Salmonella is associated with decreased mRNA expression of virulence genes invA and avrA, growth and intracellular invasion and survival. , 2009, Veterinary microbiology.

[25]  Y. Hara-Kudo,et al.  Detection of Verotoxigenic Escherichia coli O157 and O26 in food by plating methods and LAMP method: a collaborative study. , 2008, International journal of food microbiology.