title: Development and preliminary application of a quantitative PCR assay for detecting gtxA-containing Gallibacterium species in chickens.
abstract: A quantitative PCR (qPCR) assay using SYBR Green I was developed based on the published sequence of the gtxA gene from Gallibacterium anatis. This method produced reliable specificity, sensitivity, and repeatability. The detection rate of Gallibacterium in 181 clinical samples was 36.5% (66/181) by qPCR, which was superior to the detection rate of Gallibacterium-specific PCR (0/181) and an isolation and identification assay (18.2% or 33/181). No association was found between the prevalence of Gallibacterium and the age of the chickens. Gallibacterium infection was detected in one 4-day-old chicken, showing that infection can occur much earlier than the previously stated fourth week of life. Tissue sample analysis showed that Gallibacterium is mainly located in the trachea and ovaries, based on results from three groups of chicken with different health statuses. Furthermore, a titer analysis suggested that Gallibacterium loads in different organs may correlate with different clinical manifestations of disease. Thus, the qPCR assay developed in the present study is useful for identification and quantitative analysis of gtxA-containing Gallibacterium in various tissue samples from birds and for the assessment of the pathogenic mechanisms of Gallibacterium.
authors.affiliation: Institute of Poultry Diseases, Henan Agricultural University, Zhengzhou 450002, China.
authors.forename: Heping
authors.initials: H
authors.lastname: Huangfu
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
authors.forename: Xia
authors.initials: X
authors.lastname: Yang
authors.forename: Lu
authors.initials: L
authors.lastname: Chen
authors.forename: Hongtao
authors.initials: H
authors.lastname: Chang
authors.forename: Xinwei
authors.initials: X
authors.lastname: Wang
authors.forename: Qiaojing
authors.initials: Q
authors.lastname: Li
authors.forename: Huixia
authors.initials: H
authors.lastname: Yao
authors.forename: Chuanqing
authors.initials: C
authors.lastname: Wang
mesh_terms: D000367:Age Factors; D000818:Animals; D002645:Chickens; D006304:Health Status; D008969:Molecular Sequence Data; D015661:Pasteurellaceae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016871:Pasteurellaceae Infections / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D011201:Poultry Diseases / Q000453:epidemiology* / Q000382:microbiology; D015995:Prevalence; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000662:veterinary; D013045:Species Specificity; D014018:Tissue Distribution; D037521:Virulence Factors / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D037521:Virulence Factors