Novel multiplex-PCR for rapid detection of Bacillus anthracis spores present in soils of Sirajganj district in Bangladesh

Bacillus anthracis spores were isolated and detected from soil samples (n=72) using multiplex-PCR method. The bacteria were isolated and primarily identified as Bacillus anthracis using selective Polymyxin B - Lysozyme - EDTA - Thallous acetate agar. A multiplex-PCR method targeting three genes; rpoB of genome, pag of pX01 and cap of pX02 was done to confirm the isolated bacteria. Among 72 soil samples, the viable B. anthracis spores could be extracted from 14 (19.44%) samples. However, both pX01 and pX02 plasmids were harbored in 5 (6.94%) isolates. On the other hand, pX01 and pX02 was present in 8 (57.14%) and 11 (78.57%) isolates, respectively. This two-step-method was found to be easy, accurate and rapid in identification of B. anthracis spores from soil samples and to identify the toxigenic plasmids present in this bacterium. Progressive Agriculture 26:67-70, 2015

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A New Multiplex PCR Method for Rapid Detection of Genes Encoding VIM and IMP Types of Metallo Beta Lactamases

Turkiye Klinikleri Journal of Medical Sciences ◽

10.5336/medsci.2009-11518 ◽

2010 ◽

Vol 30 (4) ◽

pp. 1312-1316 ◽

Cited By ~ 2

Author(s):

Ömer KÜÇÜKBASMACI ◽

Kenan MİDİLLİ ◽

Ghassan ISSA ◽

Özlem GÜVEN ◽

Nevriye GÖNÜLLÜ

Keyword(s):

Multiplex Pcr ◽

Rapid Detection ◽

Beta Lactamases ◽

Genes Encoding ◽

Pcr Method

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Improved multiplex PCR method for the rapid detection of β-lactamase genes in Escherichia coli of animal origin

Diagnostic Microbiology and Infectious Disease ◽

10.1016/j.diagmicrobio.2006.03.005 ◽

2006 ◽

Vol 56 (1) ◽

pp. 103-106 ◽

Cited By ~ 12

Author(s):

Constança Pomba ◽

Nuno Mendonça ◽

Marta Costa ◽

Deolinda Louro ◽

Bruno Baptista ◽

...

Keyword(s):

Escherichia Coli ◽

Multiplex Pcr ◽

Rapid Detection ◽

Animal Origin ◽

Pcr Method

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Faculty Opinions recommendation of Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.726144850.793519432 ◽

2016 ◽

Author(s):

Graham Hatfull ◽

Welkin Pope

Keyword(s):

Bacillus Anthracis ◽

Rapid Detection ◽

Environmental Samples ◽

Bacillus Anthracis Spores

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Real-Time PCR Assay for Rapid Detection of Bacillus anthracis Spores in Clinical Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.40.11.4399.2002 ◽

2002 ◽

Vol 40 (11) ◽

pp. 4399-4399 ◽

Cited By ~ 35

Author(s):

L. Drago ◽

A. Lombardi ◽

E. D. Vecchi ◽

M. R. Gismondo

Keyword(s):

Bacillus Anthracis ◽

Real Time ◽

Real Time Pcr ◽

Rapid Detection ◽

Clinical Samples ◽

Pcr Assay ◽

Bacillus Anthracis Spores

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Natural Dissemination of Bacillus anthracis Spores in Northern Canada

Applied and Environmental Microbiology ◽

10.1128/aem.71.3.1610-1615.2005 ◽

2005 ◽

Vol 71 (3) ◽

pp. 1610-1615 ◽

Cited By ~ 40

Author(s):

D. C. Dragon ◽

D. E. Bader ◽

J. Mitchell ◽

N. Woollen

Keyword(s):

Bacillus Anthracis ◽

Sample Selection ◽

Soil Samples ◽

The Body ◽

Complete Coverage ◽

Phenotypic Analysis ◽

Significant Difference ◽

Bacillus Anthracis Spores ◽

Wood Buffalo National Park ◽

A Site

ABSTRACT Soil samples were collected from around fresh and year-old bison carcasses and areas not associated with known carcasses in Wood Buffalo National Park during an active anthrax outbreak in the summer of 2001. Sample selection with a grid provided the most complete coverage of a site. Soil samples were screened for viable Bacillus anthracis spores via selective culture, phenotypic analysis, and PCR. Bacillus anthracis spores were isolated from 28.4% of the samples. The highest concentrations of B. anthracis spores were found directly adjacent to fresh carcasses and invariably corresponded to locations where the soil had been saturated with body fluids escaping the carcass through either natural body orifices or holes torn by scavengers. The majority of positive samples were found within 2 m of both year-old and fresh carcasses and probably originated from scavengers churning up and spreading the body fluid-saturated soil as they fed. Trails of lesser contamination radiating from the carcasses probably resulted from spore dissemination through adhesion to scavengers and through larger scavengers dragging away disarticulated limbs. Comparison of samples from minimally scavenged and fully necropsied carcass sites revealed no statistically significant difference in the level of B. anthracis spore contamination. Therefore, the immediate area around a suspected anthrax carcass should be considered substantially contaminated regardless of the condition of the carcass.

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Intra- and inter-laboratory evaluation of an improved multiplex-PCR method for detection and typing of Salmonella

The Journal of Infection in Developing Countries ◽

10.3855/jidc.2445 ◽

2012 ◽

Vol 6 (05) ◽

pp. 443-451 ◽

Cited By ~ 1

Author(s):

Ilargi Martinez-Ballesteros ◽

Bianca Paglietti ◽

Aitor Rementeria ◽

Lorena Laorden ◽

Maria Garcia-Ricobaraza ◽

...

Keyword(s):

Developing Countries ◽

Multiplex Pcr ◽

Rapid Detection ◽

Basque Country ◽

Pcr Primers ◽

Laboratory Evaluation ◽

Kappa Index ◽

Pcr Method ◽

Sensitivity Specificity ◽

The University

Introduction: We developed and evaluated a multiplex-PCR method for rapid detection of the most common Salmonella serovars in both developed and developing countries. Additionally, the stability of the premixed reagents at high room temperature was studied. Methodology: Fifty-two Salmonella strains belonging to the collections of the University of Sassari, Italy, and to the University of the Basque Country, Spain, and a collection of a hundred blinded strains, were used to evaluate the multiplex-PCR. Primers targeting genes STY1599 and fliC were selected, and the method was evaluated both intra and inter-laboratories. Results: The inter-laboratory reproducibility was 95.92%, with a kappa index of 0.757 that indicates a substantial agreement and a high accuracy (80.81%). The sensitivity, specificity, accuracy and precision indexes for the Salmonella genus and S. Typhi targets were maximum, although the targets for Paratyphi A, Typhimurium and Enteritidis showed less accuracy. During a seven-week period, hot-start multiplex-PCR runs were performed with reagents mixed with wax to test their stability at 30ºC, and no significant variation in the patterns of amplification was observed. Conclusions: An improved multiplex-PCR for rapid detection of the most common serovars of Salmonella operable in both developed and developing countries has been designed and tested intra and inter-laboratories. Following a careful optimization protocol will not only allow the confirmation of any suspicious colony by the amplification of the Salmonella genus target, but also the preliminary adscription to the prevalent serovars. Premixed reagents with wax facilitate the throughput and stability of reagents at high room temperatures.

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