title: Detection of infectious laryngotracheitis virus by real-time PCR in naturally and experimentally infected chickens.
abstract: Infectious laryngotracheitis (ILT) is an acute, highly contagious upper-respiratory infectious disease of chickens. In this study, a real-time PCR method was developed for fast and accurate detection and quantitation of ILTV DNA of chickens experimentally infected with ILTV strain LJS09 and naturally infected chickens. The detection lower limit of the assay was 10 copies of DNA. There were no cross reactions with the DNA and RNA of infectious bursal disease virus, chicken anemia virus, reticuloendotheliosis virus, avian reovirus, Newcastle disease virus, and Marek's disease virus. The real-time PCR was reproducible as the coefficients of variation of reproducibility of the intra-assay and the inter-assay were less than 2%. The real-time PCR was used to detect the levels of the ILTV DNA in the tissues of specific pathogen free (SPF) chickens infected with ILTV at different times post infection. ILTV DNA was detected by real-time PCR in the heart, liver, spleen, lung, kidney, larynx, tongue, thymus, glandular stomach, duodenum, pancreatic gland, small intestine, large intestine, cecum, cecal tonsil, bursa of Fabricius, and brain of chickens in the infection group and the contact-exposure group. The sensitivity, specificity, and reproducibility of the ILTV real-time PCR assay revealed its suitability for detection and quantitation of ILTV in the samples from clinically and experimentally ILTV infected chickens.
authors.affiliation: Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Congcong
authors.initials: C
authors.lastname: Kong
authors.forename: Xianlan
authors.initials: X
authors.lastname: Cui
authors.forename: Hongyu
authors.initials: H
authors.lastname: Cui
authors.forename: Xingming
authors.initials: X
authors.lastname: Shi
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Zhang
authors.forename: Shunlei
authors.initials: S
authors.lastname: Hu
authors.forename: Lianwei
authors.initials: L
authors.lastname: Hao
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002645:Chickens; D003429:Cross Reactions / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D025921:Iltovirus / Q000235:genetics*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000235:genetics / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology*; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms / Q000235:genetics
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral