EVALUATION OF THE ISOLATION AND DETECTION METHODS FOR SALMONELLA SPP. FROM EGG SHELL CONTAMINATION USING MULTIPLEX PCR.

Microbial water-borne diseases still affect developing countries and are major water quality concerns throughout the world. Routine culture-based methods of identifying bacterial pathogens in water sources are laborious and time-consuming. Recently, the use of molecular techniques such as the polymerase chain reaction (PCR) has provided rapid and highly promising detection methods. In this study, we developed two multiplex PCR assays for simultaneous detection of six water-borne bacteria. Two triplex PCR protocols were developed to detect six target genes. The first protocol targets uidA (Escherichia coli), int (Shigella spp.), and gyrB (Pseudomonas aeruginosa) genes, while invA (Salmonella spp.), ompW (Vibrio cholera), and lacZ (coliforms) were amplified by the second protocol. Specificity testing was carried out for 12 reference strains. Furthermore, the applicability of the multiplex PCR assays for detection of these bacteria was investigated for 52 surface water samples. The results indicated that all primer pairs showed specificities only for their corresponding target organisms. The detection sensitivity of both multiplex PCR assays was 3 × 102 − 3 × 103 colony forming units. The developed assays represent simple and efficient diagnostic procedures for co-detection of water-borne bacteria and have the potential to provide earlier warnings of possible public health threats and more accurate surveillance of these organisms.

Download Full-text

Comparison of three human papillomavirus DNA detection methods: Next generation sequencing, multiplex-PCR and nested-PCR followed by Sanger based sequencing

Journal of Medical Virology ◽

10.1002/jmv.24413 ◽

2015 ◽

Vol 88 (5) ◽

pp. 888-894 ◽

Cited By ~ 14

Author(s):

Allex Jardim da Fonseca ◽

Renata Silva Galvão ◽

Angelica Espinosa Miranda ◽

Luiz Carlos de Lima Ferreira ◽

Zigui Chen

Keyword(s):

Human Papillomavirus ◽

Next Generation Sequencing ◽

Multiplex Pcr ◽

Nested Pcr ◽

Dna Detection ◽

Detection Methods ◽

Next Generation ◽

Papillomavirus Dna ◽

Generation Sequencing

Download Full-text

Multiplex PCR for the concurrent detection and differentiation of Salmonella spp., Salmonella Typhi and Salmonella Typhimurium

Tropical Medicine and Health ◽

10.2149/tmh.2010-20 ◽

2011 ◽

Vol 39 (1) ◽

pp. 9-15 ◽

Cited By ~ 11

Author(s):

Chai Fung Pui ◽

Woan Chwen Wong ◽

Lay Ching Chai ◽

Hai Yen Lee ◽

Ahmad Noorlis ◽

...

Keyword(s):

Salmonella Typhimurium ◽

Multiplex Pcr ◽

Salmonella Typhi ◽

Salmonella Spp

Download Full-text

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

Indian Journal of Animal Research ◽

10.18805/ijar.b-4695 ◽

2021 ◽

Author(s):

Pankaj Kumar ◽

Abhay Kumar ◽

Kamal Sarma ◽

Paresh Sharma ◽

Rashmi Rekha Kumari ◽

...

Keyword(s):

Multiplex Pcr ◽

Large Scale ◽

Parasitic Infection ◽

Epidemiological Studies ◽

Kappa Coefficient ◽

Detection Methods ◽

Diagnostic Process ◽

Specific Gene ◽

Rrna Gene ◽

Babesia Bigemina

Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in bovine blood co-infected with three of the most common hemo-parasites. Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.

Download Full-text

Fast and Efficient Mutation Detection Method Using Multiplex PCR and Cycle Sequencing

Thrombosis and Haemostasis ◽

10.1055/s-0037-1613794 ◽

2000 ◽

Vol 83 (02) ◽

pp. 244-247 ◽

Cited By ~ 11

Author(s):

J. M. Costa ◽

P. Ernault ◽

D. Vidaud ◽

M. Vidaud ◽

D. Meyer ◽

...

Keyword(s):

Multiplex Pcr ◽

Mutation Detection ◽

Detection Method ◽

Deleterious Mutation ◽

Low Cost ◽

Detection Methods ◽

Cycle Sequencing ◽

Haemophilia B ◽

Mild Haemophilia

SummaryA method using multiplex PCR followed by cycle-sequencing has been developed to detect mutations in the FIX gene. The procedure was evaluated in 45 severe or mild haemophilia B patients from 45 unrelated families. At least one deleterious mutation was identified in every haemophiliac demonstrating the efficiency of the method. Furthermore the described procedure offers many advantages compared to other screening detection methods: it is fast (less than 48 h), simple (partly automated) and of relatively low cost (it requires only one PCR).

Download Full-text

Detection of potential pathogenic aerobic bacteria from egg shell and egg contents of hen collected from poultry

Bangladesh Medical Research Council Bulletin ◽

10.3329/bmrcb.v41i2.29983 ◽

2016 ◽

Vol 41 (2) ◽

pp. 67-72

Author(s):

Jannatul Fardous ◽

S.M Shamsuzzaman

Keyword(s):

Salmonella Enteritidis ◽

Enterobacter Aerogenes ◽

Salmonella Typhi ◽

Aerobic Bacteria ◽

Salmonella Spp ◽

Providencia Rettgeri ◽

Egg Shells ◽

Egg Shell ◽

Growth Of Bacteria ◽

Hen Egg

This study was done to identify different pathogenic aerobic bacteria from egg shell and egg contents of hen. Egg shells and egg contents of 150 eggs collected from poultry were tested. Of 150 egg shells, 130 (86.67%) yielded growth of bacteria and 60 (40%) Esch. coli, 25 (16.67%) Providencia rettgeri, 5 (3.33%) Providencia alkalifaciens, 20 (13.33%) Citrobacter freundii, 10 (6.67%) Salmonella spp, 10 (6.67%) Enterobacter aerogenes were isolated. No bacteria were isolated from 150 egg contents. Total 14 (9.33%) Salmonella spp. from egg shells and 7 (4.67%) Salmonella spp. from egg contents were identified by PCR. Most of the identified serotypes were Salmonella Enteritidis (42.86% from egg shells and 71.43% from egg contents). All (100%) Salmonella Typhi and Salmonella Paratyphi A were sensitive to ciprofloxacin and ceftriaxone.

Download Full-text

Development of a multiplex PCR detection of Salmonella spp. and Shigella spp. in mussels

Letters in Applied Microbiology ◽

10.1046/j.1365-2672.2000.00797.x ◽

2000 ◽

Vol 31 (2) ◽

pp. 105-109 ◽

Cited By ~ 36

Author(s):

A. Vantarakis ◽

G. Komninou ◽

D. Venieri ◽

M. Papapetropoulou

Keyword(s):

Multiplex Pcr ◽

Pcr Detection ◽

Salmonella Spp ◽

Shigella Spp

Download Full-text

Characterization of Antibiotic Resistant Salmonella spp Isolated from Chicken Eggs of Dhaka City

Journal of Scientific Research ◽

10.3329/jsr.v3i1.6109 ◽

2010 ◽

Vol 3 (1) ◽

pp. 191 ◽

Cited By ~ 12

Author(s):

M. M. Ahmed ◽

M. M. Rahman ◽

K. R. Mahbub ◽

M. Wahiduzzaman

Keyword(s):

Antibiotic Resistance ◽

Salmonella Enterica ◽

Egg Yolk ◽

Dhaka City ◽

Salmonella Spp ◽

Antibiotic Resistant ◽

Poultry Eggs ◽

Potential Reservoir ◽

Egg Shell ◽

Chicken Eggs

In present study, the occurrence of Salmonella in local chicken eggs and their pattern of antibiotic resistance were determined. 100 egg samples collected from different locations of Dhaka city were analysed and Salmonella spp were found in 8% of the samples. Among all presumptive Salmonella isolates, 8 isolates were confirmed as Salmonella enterica subsp. salamae (4%), Salmonella enterica subsp. indica (1%), Salmonella Paratyphi-A (1%), Salmonella bongori (1%) and Salmonella Choleraesuis (1%) on the basis of serotyping and biochemical analysis. These isolates were subjected to susceptibility test against 10 antibiotic disks. All the isolates were found chloramphenicol sensitive. The highest percentage of resistance (87.5%) was found to amoxicillin and ampicillin. Resistance against erythromycin, cephalexin, doxycycline hydrochloride, ceftazidime, doxycycline and nalidixic acid was also found significant ranging from 25% to 62.5%. Salmonella isolated from egg shell surface were found more antibiotic resistant than that of egg yolk and white. The present study suggests that poultry eggs are potential reservoir of antibiotic resistant Salmonellae.Keywords: Egg; Salmonella; Characterization; Antibiotic resistance.© 2011 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.doi:10.3329/jsr.v3i1.6109 J. Sci. Res. 3 (1), 191-196 (2011)

Download Full-text