scholarly journals Development of a Two-Step Multiplex PCR Assay for Typing of Capsular Polysaccharide Synthesis Gene Clusters of Streptococcus suis

2014 ◽  
Vol 52 (5) ◽  
pp. 1714-1719 ◽  
Author(s):  
M. Okura ◽  
C. Lachance ◽  
M. Osaki ◽  
T. Sekizaki ◽  
F. Maruyama ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Capsular Polysaccharide ◽  
Streptococcus Suis ◽  
Gene Clusters ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Polysaccharide Synthesis

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 Detection and Identification ofActinobacillus pleuropneumoniae Serotype 5 by Multiplex PCR

1998 ◽  
Vol 36 (6) ◽  
pp. 1704-1710 ◽  
Author(s):  
Terry M. Lo ◽  
Christine K. Ward ◽  
Thomas J. Inzana
Keyword(s):  
Early Detection ◽  
Multiplex Pcr ◽  
Sensitivity And Specificity ◽  
Dna Sequences ◽  
Capsular Polysaccharide ◽  
Actinobacillus Pleuropneumoniae ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Detection And Identification ◽  
Serotype 5

Serotyping of Actinobacillus pleuropneumoniae is based on detection of the serotype-specific capsular antigen. However, not all isolates can be serotyped, and some may cross-react with multiple serotyping reagents. To improve sensitivity and specificity of serotyping and for early detection, a multiplex PCR assay was developed for detection of A. pleuropneumoniae and identification of serotype 5 isolates. DNA sequences specific to the conserved export and serotype-specific biosynthesis regions of the capsular polysaccharide of A. pleuropneumoniaeserotype 5 were used as primers to amplify 0.7- and 1.1-kb DNA fragments, respectively. The 0.7-kb fragment was amplified from all strains of A. pleuropneumoniae tested with the exception of serotype 4. The 0.7-kb fragment was not amplified from any heterologous species that are also common pathogens or commensals of swine. In contrast, the 1.1-kb fragment was amplified from all serotype 5 strains only. The assay was capable of amplifying DNA from less than 102 CFU. The A. pleuropneumoniaeserotype 5 capsular DNA products were readily amplified from lung tissues obtained from infected swine, although the 1.1-kb product was not amplified from some tissues stored frozen for 6 years. The multiplex PCR assay enabled us to detect A. pleuropneumoniae rapidly and to distinguish serotype 5 strains from other serotypes. The use of primers specific to the biosynthesis regions of other A. pleuropneumoniae serotypes would expand the diagnostic and epidemiologic capabilities of this assay.

scholarly journals Detection and differentiation of Burkholderia species with pathogenic potential in environmental soil samples

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245175
Author(s):  
Sujintana Janesomboon ◽  
Veerachat Muangsombut ◽  
Varintip Srinon ◽  
Chatruthai Meethai ◽  
Chayada S. Tharinjaroen ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Capsular Polysaccharide ◽  
High Sensitivity ◽  
Pcr Primers ◽  
Soil Samples ◽  
Pcr Assay ◽  
Pathogenic Potential ◽  
Specific Pcr ◽  
Phylogenetic Cluster ◽  
Multiplex Pcr Assay

The Burkholderia pseudomallei phylogenetic cluster includes B. pseudomallei, B. mallei, B. thailandensis, B. oklahomensis, B. humptydooensis and B. singularis. Regarded as the only pathogenic members of this group, B. pseudomallei and B. mallei cause the diseases melioidosis and glanders, respectively. Additionally, variant strains of B. pseudomallei and B. thailandensis exist that include the geographically restricted B. pseudomallei that express a B. mallei-like BimA protein (BPBM), and B. thailandensis that express a B. pseudomallei-like capsular polysaccharide (BTCV). To establish a PCR-based assay for the detection of pathogenic Burkholderia species or their variants, five PCR primers were designed to amplify species-specific sequences within the bimA (Burkholderia intracellular motility A) gene. Our multiplex PCR assay could distinguish pathogenic B. pseudomallei and BPBM from the non-pathogenic B. thailandensis and the BTCV strains. A second singleplex PCR successfully discriminated the BTCV from B. thailandensis. Apart from B. humptydooensis, specificity testing against other Burkholderia spp., as well as other Gram-negative and Gram-positive bacteria produced a negative result. The detection limit of the multiplex PCR in soil samples artificially spiked with known quantities of B. pseudomallei and B. thailandensis were 5 and 6 CFU/g soil, respectively. Furthermore, comparison between standard bacterial culture and the multiplex PCR to detect B. pseudomallei from 34 soil samples, collected from an endemic area of melioidosis, showed high sensitivity and specificity. This robust, sensitive, and specific PCR assay will be a useful tool for epidemiological study of B. pseudomallei and closely related members with pathogenic potential in soil.

scholarly journals Development of a multiplex PCR assay to detect the major clonal complexes of Streptococcus suis relevant to human infection

2016 ◽  
Vol 65 (5) ◽  
pp. 392-396 ◽  
Author(s):  
Rujirat Hatrongjit ◽  
Anusak Kerdsin ◽  
Marcelo Gottschalk ◽  
Shigeyuki Hamada ◽  
Kazunori Oishi ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Streptococcus Suis ◽  
Human Infection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

New approach for the identification of potentially toxigenic Corynebacterium sp. using a multiplex PCR assay

2021 ◽  
pp. 106198
Author(s):  
Sunarno ◽  
Khariri ◽  
Fauzul Muna ◽  
Kambang Sariadji ◽  
Yuni Rukminiati ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Assay ◽  
New Approach ◽  
Multiplex Pcr Assay ◽  
Corynebacterium Sp
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