Triplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis

$48.00 plus tax (Refund Policy)

Download / Buy Article:

Abstract:

Xu Y, Xu Y, Hou Q, Yang R, Zhang S. Triplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis. APMIS 2010; 118: 685–91.

A triplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis was developed. Three targets were used for amplification in a single tube: the insertion sequence IS481 and the pertussis toxin promoter region (ptxP) for B. pertussis, and the insertion sequence IS1001 for B. parapertussis. The performance of this PCR assay was evaluated in parallel in three single-target real-time PCR assays using DNA extracted from B. pertussis and B. parapertussis reference strains and nasopharyngeal swabs taken from 105 patients who had been coughing for more than 7 days. The minimum detection limit of the triplex PCR was one to five colony-forming units (CFU) of B. pertussis and 1 CFU of B. parapertussis per reaction, and the coefficients of both intra- and inter-assay variation were less than 7%. Results were available within 4 h. Of the 105 nasopharyngeal samples, seven were culture positive and 23 were PCR positive for B. pertussis. All culture-positive samples were also PCR positive. Our single-tube triplex real-time PCR assay proved to be sensitive, specific and suitable for simultaneous detection and discrimination of B. pertussis and B. parapertussis.

Keywords: Bordetella parapertussis; Bordetella pertussis; Triplex; diagnosis; real-time PCR

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1600-0463.2010.02644.x

Affiliations: 1: Department of Laboratory Medicine, The Second Hospital of Shijiazhuang City, Shijiazhuang, China 2: Department of Serum, National Institute for the Control of Pharmaceutical and Biological Products, Beijing 3: Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing

Publication date: September 1, 2010

Related content

Tools

Favourites

Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more