scholarly journals Detection and Differentiation ofListeria spp. by a Single Reaction Based on Multiplex PCR

1999 ◽  
Vol 65 (10) ◽  
pp. 4688-4692 ◽  
Author(s):  
Andreas Bubert ◽  
Inge Hein ◽  
Marcus Rauch ◽  
Angelika Lehner ◽  
ByoungSu Yoon ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Dna Sequences ◽  
Simultaneous Detection ◽  
Specific Gene ◽  
Food Hygiene ◽  
Specific Detection ◽  
Pcr Method ◽  
Species Specific ◽  
Different Sources ◽  
Single Reaction

ABSTRACT The iap gene encodes the protein p60, which is common to all Listeria species. A previous comparison of the DNA sequences indicated conserved and species-specific gene portions. Based on these comparisons, a combination consisting of only five different primers that allows the specific detection and differentiation ofListeria species with a single multiplex PCR and subsequent gel analysis was selected. One primer was derived from the conserved 3′ end and is specific for all Listeria species; the other four primers are specific for Listeria monocytogenes,L. innocua, L. grayi, or the three grouped species L. ivanovii, L. seeligeri, and L. welshimeri, respectively. The PCR method, which also enables the simultaneous detection of L. monocytogenes and L. innocua, was evaluated against conventional biotyping with 200 food hygiene-relevant Listeria strains. The results indicated the superiority of this technique. Thus, this novel type of multiplex PCR may be useful for rapid Listeria species confirmation and for identification of Listeria species for strains isolated from different sources.

A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

Food Control ◽  
2009 ◽  
Vol 20 (4) ◽  
pp. 366-370 ◽  
Author(s):  
Weibin Bai ◽  
Wentao Xu ◽  
Kunlun Huang ◽  
Yanfang Yuan ◽  
Sishuo Cao ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Species ◽  
Pcr Method

Cloth-Based Hybridization Array System for Expanded Identification of the Animal Species Origin of Derived Materials in Feeds

2007 ◽  
Vol 70 (12) ◽  
pp. 2900-2905 ◽  
Author(s):  
JOHANNA MURPHY ◽  
JENNIFER ARMOUR ◽  
BURTON W. BLAIS
Keyword(s):  
Mitochondrial Dna ◽  
Dna Sequences ◽  
Microscopic Examination ◽  
Animal Species ◽  
Specific Detection ◽  
Animal Feeds ◽  
Species Origin ◽  
Pcr Products ◽  
Species Specific ◽  
Immunoenzymatic Assay

A cloth-based hybridization array system (CHAS) previously developed for the detection of animal species for which prohibited materials have been specified (cattle, sheep, goat, elk, and deer) has been expanded to include the detection of animal species for which there are no prohibitions (pig and horse) in Canadian and American animal feeds. Animal species were identified by amplification of mitochondrial DNA sequences by PCR and subsequent hybridization of the amplicons with an array of species-specific oligonucleotide capture probes immobilized on a polyester cloth support, followed by an immunoenzymatic assay of the bound PCR products. The CHAS permitted sensitive and specific detection of meat meals from different animal species blended in a grain-based feed and should provide a useful adjunct to microscopic examination for the identification of prohibited materials in animal feeds.

Multiplex PCR Method for the Simultaneous Detection of Histamine-, Tyramine-, and Putrescine-Producing Lactic Acid Bacteria in Foods

2005 ◽  
Vol 68 (4) ◽  
pp. 874-878 ◽  
Author(s):  
ÁNGELA MARCOBAL ◽  
BLANCA de las RIVAS ◽  
M. VICTORIA MORENO-ARRIBAS ◽  
ROSARIO MUÑOZ
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Fermented Foods ◽  
Multiplex Pcr Assay ◽  
Pcr Method ◽  
Dna Fragment ◽  
Optimized Conditions ◽  
Selection Of

In a screening of primers, we have selected three pairs of primers for a multiplex PCR assay for the simultaneous detection of lactic acid bacteria (LAB) strains, which potentially produce histamine, tyramine, and putrescine on fermented foods. These primers were based on sequences from histidine, tyrosine, and ornithine decarboxylases from LAB. Under the optimized conditions, the assay yielded a 367-bp DNA fragment from histidine decarboxylases, a 924-bp fragment from tyrosine decarboxylases, and a 1,446-bp fragment from ornithine decarboxylases. When the DNAs of several target organisms were included in the same reaction, two or three corresponding amplicons of different sizes were observed. This assay was useful for the detection of amine-producing bacteria in control collection strains and in a LAB collection. No amplification was observed with DNA from nonproducing LAB strains. This article is the first describing a multiplex PCR approach for the simultaneous detection of potentially amine-producing LAB in foods. It can be easily incorporated into the routine screening for the accurate selection of starter LAB and in food control laboratories.

Development of a sensitive and specific multiplex PCR method for the simultaneous detection of chicken, duck and goose DNA in meat products

Meat Science ◽  
2015 ◽  
Vol 101 ◽  
pp. 90-94 ◽  
Author(s):  
Bo Hou ◽  
Xianrong Meng ◽  
Liyuan Zhang ◽  
Jinyue Guo ◽  
Shaowen Li ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Products ◽  
Pcr Method

scholarly journals Standardization of a rapid quadruplex PCR method for the simultaneous detection of bovine, buffalo, Salmonella spp., and Listeria monocytogenes DNA in milk

2021 ◽  
Vol 73 (4) ◽  
pp. 781-790
Author(s):  
J.S. Lima ◽  
A.P.P.O. Sampaio ◽  
M.C.S. Dufossé ◽  
A.M.B.P. Rosa ◽  
P.F.M. Sousa ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Dna Sequences ◽  
Performance Optimization ◽  
Genetic Material ◽  
Simultaneous Detection ◽  
Buffalo Milk ◽  
Salmonella Spp ◽  
Initial Inoculum ◽  
Target Dna ◽  
Pcr Method

ABSTRACT The objective of the present study was to Standardize a Polymerase Chain Reaction (PCR) protocol for the authentication of bovine and buffalo milk, and to detect the presence of Salmonella spp. and Listeria monocytogenes. For this, the target DNA was extracted, mixed, and subjected to a PCR assay. Milk samples were defrauded and experimentally contaminated with microorganisms to assess the detection of target DNA at different times of cultivation, bacterial titers, and concentration of genetic material. In addition, the protocol was tested with DNA extracted directly from food, without a pre-enrichment step. The proposed quadruplex PCR showed good accuracy in identifying target DNA sequences. It was possible to simultaneously identify all DNA sequences at the time of inoculation (0h), when the samples were contaminated with 2 CFU/250mL and with 6h of culture when the initial inoculum was 1 CFU/250mL. It was also possible to directly detect DNA sequences from the food when it was inoculated with 3 CFU/mL bacteria. Thus, the proposed methodology showed satisfactory performance, optimization of the analysis time, and a potential for the detection of microorganisms at low titers, which can be used for the detection of fraud and contamination.

scholarly journals Development and application of multiplex PCR method for simultaneous detection of seven viruses in ducks

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Ming Yao ◽  
Xiyu Zhang ◽  
Yunfei Gao ◽  
Suquan Song ◽  
Danning Xu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Pcr Method

A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

2014 ◽  
Vol 59 (1) ◽  
Author(s):  
Junlong Liu ◽  
Guiquan Guan ◽  
Aihong Liu ◽  
Youquan Li ◽  
Hong Yin ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Internal Transcribed Spacers ◽  
Babesia Bovis ◽  
Babesia Bigemina ◽  
Pcr Assay ◽  
Blood Samples ◽  
Multiplex Pcr Assay ◽  
Pcr Method

AbstractIn this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

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