High-Resolution Melting PCR as Rapid Genotyping Tool for Brucella Species
TLDR
A high-resolution melting (HRM) method for the rapid screening of DNA and direct assignment into one of the 12 species of the Brucella genus based on single nucleotide polymorphisms (SNPs) based on whole genome SNP analysis based on 988 genomes.Abstract:
Brucella sp. are the causative agents of brucellosis. One of the main characteristics of the Brucella genus concerns its very high genetic homogeneity. To date, classical bacteriology typing is still considered as the gold standard assay for direct diagnosis of Brucella. Molecular approaches are routinely used for the identification of Brucella at the genus level. However, genotyping is more complex, and to date, no method exists to quickly assign a strain into species and biovar levels, and new approaches are required. Next generation sequencing (NGS) opened a new era into the diagnosis of bacterial diseases. In this study, we designed a high-resolution melting (HRM) method for the rapid screening of DNA and direct assignment into one of the 12 species of the Brucella genus. This method is based on 17 relevant single nucleotide polymorphisms (SNPs), identified and selected from a whole genome SNP (wgSNP) analysis based on 988 genomes (complete and drafts). These markers were tested against the collection of the European Reference Laboratory (EU-RL) for brucellosis (1440 DNAs extracted from Brucella strains). The results confirmed the reliability of the panel of 17 SNP markers, allowing the differentiation of each species of Brucella together with biovars 1, 2, and 3 of B. suis and vaccine strain Rev1 (B. melitensis) within 3 h, which is a considerable gain of time for brucellosis diagnosis. Therefore, this genotyping tool provides a new and quick alternative for Brucella identification based on SNPs with the HRM-PCR assay.read more
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Journal ArticleDOI
Assays for Identification and Differentiation of Brucella Species: A Review
Berzhan K. Kurmanov,Diansy Zincke,Wanwen Su,Ted L. Hadfield,Alim Aikimbayev,Talgat Karibayev,M. Berdikulov,Mukhit Orynbayev,Mikeljon P. Nikolich,Jason K. Blackburn +9 more
TL;DR: Molecular techniques for confirming and identifying specific Brucella species are summarized and recommendations for selecting the appropriate methods based on need, sensitivity, and laboratory capabilities/technology are provided.
Journal ArticleDOI
A Combination of MALDI-TOF MS Proteomics and Species-Unique Biomarkers' Discovery for Rapid Screening of Brucellosis.
TL;DR: The presented MALDI-TOF MS proteomics analyses, coupled with special usage of bioinformatics, could be used as a cost-efficient strategy for the diagnostics of brucellosis and introduce a reliable identification protocol for species of dangerous bacteria.
Journal ArticleDOI
First record of the human infection of Brucella melitensis in Kyrgyzstan: evidence from whole-genome sequencing-based analysis
K. Kydyshov,N.T. Usenbaev,Stalbek Berdiev,Aigul K. Dzhaparova,A. Abidova,Nuraiym Kebekbaeva,Murat Abdyraev,Gamal Wareth,Hanka Brangsch,Falk Melzer,Heinrich Neubauer,Mathias W. Pletz +11 more
TL;DR: In this article , the Brucella strain B. melitensis was identified as a causative agent of human Brucellosis in Kyrgyzstan and different lineages could be identified.
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Multiplex polymerase spiral reaction for simultaneous detection of Salmonella typhimurium and Staphylococcus aureus.
Caihong Yin,Bo Pang,Yanzhi Huang,Jinhua Li,Tingyu Meng,Mengfan Zhang,Liang Zhang,Yan-Zheng Gao,Xiuling Song +8 more
TL;DR: In this article , a method for the simultaneous determination of S. typhimurium and S. aureus based on multiplex polymerase spiral reaction (m-PSR) and melting curve analysis was developed.
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Multiplex high-resolution melting assay for simultaneous detection of five key bacterial pathogens in urinary tract infections: A pilot study
Hossein Kafi,Mohammad Emaneini,Shahnaz Halimi,Hossein Ali Rahdar,Fereshteh Jabalameli,Reza Beigverdi +5 more
TL;DR: In this article , the authors developed a multiplex high-resolution melting assay (MHRM) for the simultaneous detection of five common bacterial pathogens (Escherichia coli, Klebsiella pneumoniae, Staphylococcus saprophyticus, Enterococcus faecalis, and group B streptococci (GBS)) directly from urine samples.
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