title: A nested multiplex polymerase chain reaction assay for the differential identification of three zooanthroponotic chlamydial strains in porcine swab samples.
abstract: Porcine chlamydial infection is an enzootic infectious disease caused by multiple members of the family Chlamydiaceae (e.g. Chlamydophila abortus, Chlamydia suis, and Chlamydophila pneumoniae). Rapid and accurate differentiation of these pathogens is critical in the control and prevention of disease. The aim of the current study was to develop a nested multiplex polymerase chain reaction (nmPCR) assay to simultaneously detect the 3 chlamydial pathogens in clinical samples. In the first round of the nmPCR, 1 pair of family-specific primers were used to amplify the 1,100 base pair (bp) fragment of chlamydial ompA gene. In the second round of the nmPCR, 4 inner primers were designed for Ch. abortus, C. suis, and Ch. pneumoniae. Each pathogen produced a speciﬁc amplicon with a size of 340 bp, 526 bp, and 267 bp respectively. The assay was sensitive and speciﬁc for detecting target pathogens in both cell cultures and clinical specimens. The results, incorporated with the improved rapid DNA extraction protocol, suggest that the nmPCR could be a promising assay for differential identification of different chlamydial strains in pigs.
authors: Key Laboratory of Biorheology Science and Technology Under Ministry of Education, Bio-Engineering College, Chongqing University, Chongqing 400030, China.
authors: Yingguo
authors: Y
authors: Li
authors: Yu
authors: Y
authors: Wang
authors: Fuping
authors: F
authors: Nie
authors: Jinwen
authors: J
authors: Xiao
authors: Guoming
authors: G
authors: Wang
authors: Ling
authors: L
authors: Yuan
authors: Zhengguo
authors: Z
authors: Li
mesh_terms: D000818:Animals; D001483:Base Sequence; D002693:Chlamydiaceae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002694:Chlamydiaceae Infections / Q000382:microbiology / Q000662:veterinary*; D004269:DNA, Bacterial / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology*; D015047:Zoonoses
chemical_list: D004269:DNA, Bacterial