scholarly journals Application of PCR technique in diagnosis of four respiratory pathogenic bacteria in pigs at the slaughterhouse

2019 ◽  
Vol 18 (03) ◽  
pp. 35-40
Author(s):  
Han M. Ly
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Pathogenic Bacteria ◽  
Pasteurella Multocida ◽  
Bordetella Bronchiseptica ◽  
Ho Chi Minh City ◽  
Haemophilus Parasuis ◽  
Chain Reaction ◽  
Injured Lung ◽  
Polymerase Chain

The polymerase chain reaction (PCR) for Actinobacillus pleuropneumonia (App), Haemophilus parasuis (Hps), Pasteurella multocida (Pm) and Bordetella bronchiseptica (Bb) were performed in pure colonies isolated from 114 lung specimens with lesions collected from the Vissan slaughterhouse in Ho Chi Minh City from July 2018 to May 2019. The aim of the experiment was to identify the four respiratory pathogenic bacteria in pigs at slaughterhouse by using PCR technique. The criteria for evaluating the results included the proportion of positive samples with multiplex PCR and percentage of samples co-infected with 2, 3, and 4 bacteria. Among a total of 114 injured lung samples, 21% of the samples was positive to at least one of the four bacteria, 3 samples (2.63%) were positive for App, 2 samples (1.75%) were positive for Hps, 7 samples (6.14%) were for Pm, and 12 lungs (10.53%) were positive for Bb. One sample (0.88%) was found co-infected with Pm and Hps.

scholarly journals Molecular identification by multiplex polymerase chain reaction of Pasteurella multocida in cattle and buffaloes in Baghdad

2014 ◽  
Vol 38 (1) ◽  
pp. 99-106
Author(s):  
Ihab G. M. AL-Shemmari
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Pasteurella Multocida ◽  
Type B ◽  
Chain Reaction ◽  
Blood Samples ◽  
Nasal Swabs ◽  
Polymerase Chain ◽  
B 31

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .

Diagnostic Value of Multiplex Polymerase Chain Reaction in Detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from Sepsis in Pediatrics

2021 ◽  
Vol 15 ◽  
Author(s):  
Sara Galeb ◽  
Maysaa El Sayed Zaki ◽  
Raghdaa Shrief ◽  
Rasha Hassan ◽  
Mohamed Anies
Keyword(s):  
Polymerase Chain Reaction ◽  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Stenotrophomonas Maltophilia ◽  
Multiplex Polymerase Chain Reaction ◽  
Diagnostic Value ◽  
Blood Cultures ◽  
Chain Reaction ◽  
Polymerase Chain

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. Objectives: The present study aimed to evaluate the multiplex Polymerase Chain Reaction (PCR) to detect Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children, with hospital-acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR, with primers specific to the 3 tested bacteria. Results: Multiplex PCR was positive in 96% of isolates, and 4 isolates had negative results. False positive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa, and 27 isolates of Stenotrophomonas maltophilia. Compared to the biochemical identification, the diagnostic value of the multiplex PCR revealed 96.04% sensitivity, 96.9% specificity, 97.00%, positive predictive value, 96.00% negative predictive value, and 96.50% accuracy. Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific, and accurate. The accuracy differs according to the organisms, with 100% accuracy for Acinetobacter baumannii.

Development of a Multiplex Polymerase Chain Reaction Assay for Detection and Differentiation of Staphylococcus aureus in Dairy Products†

2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Dairy Products ◽  
Multiplex Polymerase Chain Reaction ◽  
Skim Milk ◽  
Cheddar Cheese ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.

scholarly journals Prevalence and diversity of Arcobacter spp. in retail chicken meat in Turkey

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Celenk Molva ◽  
Halil Ibrahim Atabay
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Chicken Meat ◽  
Species Level ◽  
Waterborne Pathogens ◽  
Chain Reaction ◽  
Retail Markets ◽  
Specific Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Retail Chicken Meat

Arcobacters are food and waterborne pathogens associated with human and animal infections. The objective of the present study was to investigate the prevalence and diversity of <em>Arcobacter</em> spp. in commercially sold chicken meat in İzmir region of Turkey. For this purpose, 100 samples including legs (n=40), 17 chicken quarters (n=17), drumstickers (n=16), breasts (n=11), wings (n=10), and carcasses (n=6) were collected from different retail markets. A total of 65 isolates were confirmed as <em>Arcobacter</em> spp. from 55 samples by genus-specific polymerase chain reaction (PCR). The prevalence of <em>Arcobacter</em> spp. was 32.5, 81.3, 64.7, 72.7, 83.3, and 50% for legs, drumstickers, chicken quarters, breasts, carcasses and wings, respectively. Based on the multiplex-PCR, most of the isolates were identified as <em>A. butzleri</em> (n=45, 80%), followed by <em>A. cryaerophilus</em> (n=2, 3.6%), <em>A. skirrowii</em> (n=1, 1.8%) and 17 isolates (30.9%) could not be identified at the species level.

THE DETECTION OF THE GENUS LISTERIA PATHOGENIC BACTERIA BY REAL-TIME POLYMERASE CHAIN REACTION

2020 ◽  
pp. 117-125
Author(s):  
Elena V. Sokolova ◽  
Ekaterina K. Psareva ◽  
Irina Yu. Egorova ◽  
Pavel A. Zhurilov ◽  
Evgeny A. Potemkin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Pathogenic Bacteria ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Application of a Multiplex Polymerase Chain Reaction Assay for the Simultaneous Confirmation of Listeria monocytogenes and Other Listeria Species in Turkey Sample Surveillance†

2002 ◽  
Vol 65 (5) ◽  
pp. 780-785 ◽  
Author(s):  
IRENE V. WESLEY ◽  
KAREN M. HARMON ◽  
JAMES S. DICKSON ◽  
ANN RAMOS SCHWARTZ
Keyword(s):  
Polymerase Chain Reaction ◽  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Listeria Species ◽  
Multiplex Pcr Assay ◽  
Polymerase Chain

A multiplex polymerase chain reaction was developed to simultaneously identify Listeria monocytogenes and species of the genus Listeria. Two sets of primers were used, with the first amplifying a 938-bp region of the 16S rRNA gene that is highly conserved in all Listeria species and the second amplifying a 174-bp region of the listeriolysin (hlyA) gene of L. monocytogenes. Thus, isolates of Listeria spp. yield a single 938-bp product, whereas L. monocytogenes isolates yield both the 938-bp product and a 174-bp product. The specificity of the assay was verified with all six Listeria species and 11 serotypes of L. monocytogenes, as well as nonrelated bacteria. The multiplex PCR assay was used to determine the incidence of Listeria spp., especially L. monocytogenes, in mechanically separated turkey samples (n = 150 samples). L. monocytogenes strains were selected by using the University of Vermont two-step enrichment protocol and plating to selective Palcam agar. The multiplex PCR assay was used for verification of presumptive Listeria colonies. Approximately 38% of mechanically separated turkey samples (57 of 150) yielded L. monocytogenes; an additional 18% of these samples (27 of 150) harbored other Listeria spp. Fifty-one percent (29 of 57) of the L. monocytogenes isolates were of serogroup 1, 44% (25 of 57) were of serogroup 4, and 2% (1 of 57) were assigned to serogroups other than 1 and 4.

scholarly journals DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL

2014 ◽  
Vol 56 (5) ◽  
pp. 391-395 ◽  
Author(s):  
Herintha Coeto Neitzke-Abreu ◽  
Kárin Rosi Reinhold-Castro ◽  
Mateus Sabaini Venazzi ◽  
Regiane Bertin de Lima Scodro ◽  
Alessandra de Cassia Dias ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Natural Infection ◽  
Epidemiological Studies ◽  
Leishmania Infection ◽  
Southern Brazil ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sensitivity Specificity

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

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