Development of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification ofCampylobacter jejuni,Campylobacter coliandCampylobacter fetus

A multiplex PCR assay was developed by using primers to the fiber gene that could differentiate human adenovirus (Ad) species A through F in a single amplification reaction. The assay correctly identified the species of all 49 recognized Ad prototype strains as well as 180 geographically and temporally diverse Ad field isolates. Ad serotype 6 (Ad6) (species C), Ad16 (species B), Ad31 (species A), and Ad40 and Ad41 (species F) could also be distinguished by amplicon size within each respective species. In comparison, a previously described Ad species-specific multiplex PCR assay that used primers to the Ad hexon gene gave equivocal results with several serotypes of species B, whereas our multiplex assay amplified all species B serotypes equally well. Our multiplex PCR assay will permit rapid, accurate, and cost-effective classification of Ad isolates.

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A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Detection of Campylobacter spp. in Stool Specimens and Comparison with Culture Method

Journal of Veterinary Medical Science ◽

10.1292/jvms.11-0574 ◽

2012 ◽

Vol 74 (7) ◽

pp. 857-862 ◽

Cited By ~ 7

Author(s):

Sachi SHIRAMARU ◽

Masahiro ASAKURA ◽

Haruna INOUE ◽

Akira NAGITA ◽

Akio MATSUHISA ◽

...

Keyword(s):

Multiplex Pcr ◽

Culture Method ◽

Toxin Gene ◽

Cytolethal Distending Toxin ◽

Pcr Assay ◽

Multiplex Pcr Assay ◽

Stool Specimens ◽

Campylobacter Spp

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Multiplex PCR Assay for Detection of Vibrio vulnificus Biotype 2 and Simultaneous Discrimination of Serovar E Strains

Applied and Environmental Microbiology ◽

10.1128/aem.02320-06 ◽

2007 ◽

Vol 73 (6) ◽

pp. 2029-2032 ◽

Cited By ~ 21

Author(s):

Eva Sanjuán ◽

Carmen Amaro

Keyword(s):

Multiplex Pcr ◽

Vibrio Vulnificus ◽

Simultaneous Discrimination ◽

Fish Pathogen ◽

Pcr Assay ◽

Culture Collections ◽

Multiplex Pcr Assay ◽

Detection And Identification ◽

Field Samples ◽

Zoonotic Strains

ABSTRACT In the present work we develop a multiplex PCR assay for the detection and identification of the fish pathogen Vibrio vulnificus biotype 2 with discriminating potential for zoonotic strains (serovar E). The PCR assay allowed the identification of two new biotype 2 serovar E human isolates from culture collections. Finally, the multiplex was successfully applied to both diagnosis and carrier detection in field samples.

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cpn60 Gene-Based Multiplex-PCR Assay for Simultaneous Identification of Streptococcal Species

Acta Veterinaria Brno ◽

10.2754/avb200675020235 ◽

2006 ◽

Vol 75 (2) ◽

pp. 235-240 ◽

Cited By ~ 7

Author(s):

A. Dmitriev ◽

M. Bhide ◽

I. Mikula

Keyword(s):

Multiplex Pcr ◽

Streptococcus Agalactiae ◽

Streptococcus Dysgalactiae ◽

Pcr Assay ◽

Streptococcus Uberis ◽

Multiplex Pcr Assay ◽

Streptococcal Species ◽

Species Specific ◽

Simultaneous Identification

The cpn60 genes of Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis were sequenced and a certain polymorphism of cpn60 genes was revealed. Presumable species-specific pairs of primers were designed and their specificity was confirmed by PCR. Based on these data, the cpn60 gene-based multiplex-PCR assay was developed. It was found to be effective for simultaneous identification of S. agalactiae, S. dysgalactiae and S. uberis strains.

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Development of a multiplex PCR assay for the detection of key genes associated with Pasteurella multocida subspecies

Journal of Veterinary Diagnostic Investigation ◽

10.1177/10406387211063438 ◽

2021 ◽

pp. 104063872110634

Author(s):

Barbara Ujvári ◽

Hubert Gantelet ◽

Tibor Magyar

Keyword(s):

Multiplex Pcr ◽

Pasteurella Multocida ◽

Rrna Gene ◽

Pcr Assay ◽

Biochemical Tests ◽

Specific Primers ◽

Multiplex Pcr Assay ◽

Subspecies Level ◽

Species Specific ◽

Reference Strains

The ability to distinguish among the subspecies of Pasteurella multocida isolates is important epidemiologically; however, classification at the subspecies level based on the results of conventional biochemical tests (fermentation of sorbitol and dulcitol) is reportedly not accurate in all cases. Therefore, we developed a rapid, multiplex PCR assay to differentiate among the 3 subspecies of P. multocida. The PCR assay includes the P. multocida species–specific primers KMT1SP6 and KMT1T7 as an internal amplification control, with a newly designed gatD (galactitol-1-phosphate-5-dehydrogenase)-specific primer pair (unique for subsp. gallicida), and primers targeting a 16S rRNA gene region specific for subsp. septica. The subspecies specificity of the PCR was demonstrated by applying the test to a collection of 70 P. multocida isolates, including the Heddleston serovar reference strains; all isolates and strains were assigned correctly. The PCR assay is a sensitive, specific, and highly effective method for the identification of P. multocida subspecies, and an alternative to biochemical test–based differentiation. A possible relationship was noticed between P. multocida subspecies and lipopolysaccharide (LPS) genotype; all but one of the subsp. gallicida strains were isolated only from avian hosts and represented L1 LPS genotype. Subsp. multocida and subsp. septica isolates were classified into 5 and 4 different LPS genotypes, respectively, of which L3 was the only LPS genotype shared between these 2 subspecies.

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