scholarly journals Advanced Multiplex PCR Assay for Differentiation of Brucella Species

2011 ◽  
Vol 77 (18) ◽  
pp. 6726-6728 ◽  
Author(s):  
Sung-Il Kang ◽  
Moon Her ◽  
Jong Wan Kim ◽  
Ji-Yeon Kim ◽  
Kyung Yuk Ko ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Brucella Species ◽  
Pcr Assay ◽  
Content Type ◽  
Multiplex Pcr Assay ◽  
Brucella Canis ◽  
Brucella Suis ◽  
Primer Sets

ABSTRACTTwo new primer sets of a 766- and a 344-bp fragment were introduced into the conventional Bruce-ladder PCR assay. This novel multiplex PCR assay rapidly and concisely discriminatesBrucella canisandBrucella microtifromBrucella suisstrains and also may differentiate all of the 10Brucellaspecies.

scholarly journals New Bruce-ladder multiplex PCR assay for the biovar typing of Brucella suis and the discrimination of Brucella suis and Brucella canis

2011 ◽  
Vol 154 (1-2) ◽  
pp. 152-155 ◽  
Author(s):  
Ignacio López-Goñi ◽  
David García-Yoldi ◽  
Clara M. Marín ◽  
María J. de Miguel ◽  
Elías Barquero-Calvo ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Brucella Canis ◽  
Brucella Suis

Development of a multiplex PCR assay for polymorphism analysis of Brucella suis biovars causing brucellosis in swine

2006 ◽  
Vol 115 (1-3) ◽  
pp. 269-277 ◽  
Author(s):  
L FERRAOBECK ◽  
R CARDOSO ◽  
P MUNOZ ◽  
M DEMIGUEL ◽  
D ALBERT ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Polymorphism Analysis ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Brucella Suis

scholarly journals Evaluation of a Multiplex PCR Assay (Bruce-ladder) for Molecular Typing of All Brucella Species, Including the Vaccine Strains

2008 ◽  
Vol 46 (10) ◽  
pp. 3484-3487 ◽  
Author(s):  
I. Lopez-Goni ◽  
D. Garcia-Yoldi ◽  
C. M. Marin ◽  
M. J. de Miguel ◽  
P. M. Munoz ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Molecular Typing ◽  
Brucella Species ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Multiplex PCR Assay for the Identification and Differentiation of all Brucella Species and the Vaccine Strains Brucella abortus S19 and RB51 and Brucella melitensis Rev1

2006 ◽  
Vol 52 (4) ◽  
pp. 779-781 ◽  
Author(s):  
David García-Yoldi ◽  
Clara M Marín ◽  
María J de Miguel ◽  
Pilar M Muñoz ◽  
José L Vizmanos ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Brucella Abortus ◽  
Brucella Melitensis ◽  
Brucella Species ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Differentiating Botulinum Neurotoxin-Producing Clostridia with a Simple, Multiplex PCR Assay

2017 ◽  
Vol 83 (18) ◽  
Author(s):  
Charles H. D. Williamson ◽  
Adam J. Vazquez ◽  
Karen Hill ◽  
Theresa J. Smith ◽  
Roxanne Nottingham ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Gene Cluster ◽  
Botulinum Neurotoxin ◽  
Comparative Genomic ◽  
Pcr Assay ◽  
Content Type ◽  
Multiplex Pcr Assay ◽  
Group I ◽  
Pcr Assays ◽  
Group Ii

ABSTRACT Diverse members of the genus Clostridium produce botulinum neurotoxins (BoNTs), which cause a flaccid paralysis known as botulism. While multiple species of clostridia produce BoNTs, the majority of human botulism cases have been attributed to Clostridium botulinum groups I and II. Recent comparative genomic studies have demonstrated the genomic diversity within these BoNT-producing species. This report introduces a multiplex PCR assay for differentiating members of C. botulinum group I, C. sporogenes, and two major subgroups within C. botulinum group II. Coding region sequences unique to each of the four species/subgroups were identified by in silico analyses of thousands of genome assemblies, and PCR primers were designed to amplify each marker. The resulting multiplex PCR assay correctly assigned 41 tested isolates to the appropriate species or subgroup. A separate PCR assay to determine the presence of the ntnh gene (a gene associated with the botulinum neurotoxin gene cluster) was developed and validated. The ntnh gene PCR assay provides information about the presence or absence of the botulinum neurotoxin gene cluster and the type of gene cluster present (ha positive [ha +] or orfX +). The increased availability of whole-genome sequence data and comparative genomic tools enabled the design of these assays, which provide valuable information for characterizing BoNT-producing clostridia. The PCR assays are rapid, inexpensive tests that can be applied to a variety of sample types to assign isolates to species/subgroups and to detect clostridia with botulinum neurotoxin gene (bont) clusters. IMPORTANCE Diverse clostridia produce the botulinum neurotoxin, one of the most potent known neurotoxins. In this study, a multiplex PCR assay was developed to differentiate clostridia that are most commonly isolated in connection with human botulism cases: C. botulinum group I, C. sporogenes, and two major subgroups within C. botulinum group II. Since BoNT-producing and nontoxigenic isolates can be found in each species, a PCR assay to determine the presence of the ntnh gene, which is a universally present component of bont gene clusters, and to provide information about the type (ha + or orfX +) of bont gene cluster present in a sample was also developed. The PCR assays provide simple, rapid, and inexpensive tools for screening uncharacterized isolates from clinical or environmental samples. The information provided by these assays can inform epidemiological studies, aid with identifying mixtures of isolates and unknown isolates in culture collections, and confirm the presence of bacteria of interest.

scholarly journals Examining the Possibility of Detecting Brucella Canis from Tissue Samples Using Bruce-Ladder Multiplex PCR Assay

2017 ◽  
Vol 67 (4) ◽  
pp. 551-561
Author(s):  
Nataša Stević ◽  
Dušan Mišić ◽  
Danica Bogunović ◽  
Kazimir Matović ◽  
Miroslav Valčić ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Reproductive Organs ◽  
Pcr Assay ◽  
Tissue Samples ◽  
Multiplex Pcr Assay ◽  
Brucella Canis ◽  
First Choice ◽  
Two Samples ◽  
Agglutination Method ◽  
Canine Brucellosis

AbstractThe goal of this study was to compare the results of serological and conventional bacteriological methods with the results obtained using multiplex PCR Bruce-ladder assay. Based on the obtained results, the usability of the assay was assessed in regard to rapid diagnosis of canine brucellosis directly from the samples of reproductive organs of infected dogs. Out of 225 blood samples, 33 (14.67%) had a positive agglutination reaction. In this study, out of the 225 assayed reproductive organs of dogs, B. canis was isolated from 3 samples (1.33%), while the PCR Bruce-ladder assay detected two positive samples (0.88%). Two dogs from which B. canis was isolated, an antibody titer of 1/200 was established in blood serums, and third dog from which B. canis was isolated was negative using the tube agglutination test. From a total of 225 assayed organ samples, a positive PCR reaction was obtained from two samples. The obtained results show that the tube agglutination method remains the first choice for the detection of dogs infected with B. canis. In addition, whenever possible, it is necessary to try isolation. It is desirable to attempt the detection of B. canis in tissues using PCR, but the results may not be treated as definitive and reliable.

scholarly journals Multiplex PCR Assay Targeting a Diguanylate Cyclase-Encoding Gene,cgcA, To Differentiate Species within the Genus Cronobacter

2012 ◽  
Vol 79 (2) ◽  
pp. 734-737 ◽  
Author(s):  
L. Carter ◽  
L. A. Lindsey ◽  
C. J. Grim ◽  
V. Sathyamoorthy ◽  
K. G. Jarvis ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diguanylate Cyclase ◽  
Pcr Assay ◽  
Content Type ◽  
Multiplexed Pcr ◽  
Multiplex Pcr Assay ◽  
Food Borne ◽  
Encoding Gene

ABSTRACTIn a comparison to the widely usedCronobacter rpoBPCR assay, a highly specific multiplexed PCR assay based oncgcA, a diguanylate cyclase gene, that identified all of the targeted six species among 305Cronobacterisolates was designed. This assay will be a valuable tool for identifying suspectedCronobacterisolates from food-borne investigations.

scholarly journals Multiplex PCR for Identification of Two Capsular Types in Epidemic KPC-Producing Klebsiella pneumoniae Sequence Type 258 Strains

2014 ◽  
Vol 58 (7) ◽  
pp. 4196-4199 ◽  
Author(s):  
Liang Chen ◽  
Kalyan D. Chavda ◽  
Jacqueline Findlay ◽  
Gisele Peirano ◽  
Katie Hopkins ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Multiplex Pcr ◽  
Capsular Polysaccharide ◽  
Sequence Type ◽  
Pcr Assay ◽  
Content Type ◽  
Multiplex Pcr Assay ◽  
Polysaccharide Synthesis ◽  
Sequence Types

ABSTRACTWe developed a multiplex PCR assay capable of identifying two capsular polysaccharide synthesis sequence types (sequence type 258 [ST258]cps-1andcps-2) in epidemicKlebsiella pneumoniaeST258 strains. The assay performed with excellent sensitivity (100%) and specificity (100%) for identifyingcpstypes in 60 ST258K. pneumoniaesequenced isolates. The screening of 419 ST258 clonal isolates revealed a significant association betweencpstype andK. pneumoniaecarbapenemase (KPC) variant:cps-1is largely associated with KPC-2, whilecps-2is primarily associated with KPC-3.

scholarly journals Development of a One-Step Multiplex PCR Assay for Differential Detection of Major Mycobacterium Species

2017 ◽  
Vol 55 (9) ◽  
pp. 2736-2751 ◽  
Author(s):  
Hansong Chae ◽  
Seung Jung Han ◽  
Su-Young Kim ◽  
Chang-Seok Ki ◽  
Hee Jae Huh ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Beijing Genotype ◽  
Analytical Sensitivity ◽  
Pcr Assay ◽  
Accurate Identification ◽  
Reliable Technique ◽  
Content Type ◽  
Multiplex Pcr Assay ◽  
One Step ◽  
Reference Strains

ABSTRACTThe prevalence of tuberculosis continues to be high, and nontuberculous mycobacterial (NTM) infection has also emerged worldwide. Moreover, differential and accurate identification of mycobacteria to the species or subspecies level is an unmet clinical need. Here, we developed a one-step multiplex PCR assay using whole-genome analysis and bioinformatics to identify novel molecular targets. The aims of this assay were to (i) discriminate between theMycobacterium tuberculosiscomplex (MTBC) and NTM usingrv0577or RD750, (ii) differentiateM. tuberculosis(M. tuberculosis) from MTBC using RD9, (iii) selectively identify the widespreadM. tuberculosisBeijing genotype by targetingmtbk_20680, and (iv) simultaneously detect five clinically important NTM (M. avium,M. intracellulare,M. abscessus,M. massiliense, andM. kansasii) by targeting IS1311, DT1,mass_3210, andmkan_rs12360. An initial evaluation of the multiplex PCR assay using reference strains demonstrated 100% specificity for the targetedMycobacteriumspecies. Analytical sensitivity ranged from 1 to 10 pg for extracted DNA and was 103and 104CFU for pure cultures and nonhomogenized artificial sputum cultures, respectively, of the targeted species. The accuracy of the multiplex PCR assay was further evaluated using 55 reference strains and 94 mycobacterial clinical isolates. Spoligotyping, multilocus sequence analysis, and a commercial real-time PCR assay were employed as standard assays to evaluate the multiplex PCR assay with clinicalM. tuberculosisand NTM isolates. The PCR assay displayed 100% identification agreement with the standard assays. Our multiplex PCR assay is a simple, convenient, and reliable technique for differential identification of MTBC,M. tuberculosis,M. tuberculosisBeijing genotype, and major NTM species.

A Collagenase-Targeted Multiplex PCR Assay for Identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus

2005 ◽  
Vol 68 (1) ◽  
pp. 150-153 ◽  
Author(s):  
ANGELA DI PINTO ◽  
GIUSEPPINA CICCARESE ◽  
GIUSEPPINA TANTILLO ◽  
DOMENICO CATALANO ◽  
VITO TONY FORTE
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Vibrio Parahaemolyticus ◽  
Food Samples ◽  
Vibrio Alginolyticus ◽  
Pcr Assay ◽  
Molecular Approaches ◽  
Multiplex Pcr Assay ◽  
Primer Sets ◽  
Species Specific

A multiplex PCR assay using three collagenase-targeted primer pairs for the species-specific detection of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus was developed. The results highlight the species specificity of the three primer sets designed. Because of the increasing importance of Vibrio spp. in human foodborne diseases, molecular approaches for routine microbial screening and monitoring of clinical, environmental, and food samples also have become more important. The results of this study indicate that the gene coding for collagenase should be used as an alternative molecular target to discriminate among the three Vibrio species.

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