Establishment and Application of SYBR Green Ⅰ Real-Time Fluorescent Quantitative PCR Method for Chicken Infectious Laryngotracheitis Virus
Zhang Yuxia
Dong Wenwen
Yuan Xiaoyuan
Meng Kai
Xu Huaiying
Abstract：The chicken infectious laryngotracheitis virus(ILTV)has been prevalent in layer in the past,but it has been continuously isolated in breeders and broilers in recent years,indicating an expanding risk of virus infection spectrum.Therefore,it is necessary to strengthen the monitoring of ILTV.To establish an effi-cient and sensitive ILTV detection method,a pair of specific primers were designed based on the TK gene of ILTV,and then the gene was amplified and the pMD18-T-TK plasmid standard was constructed.A SYBR Green Ⅰ real-time fluorescent quantitative PCR method and standard curve were established.The specificity,sensitivity,and repeatability of the method were tested,and the ILTV suspected cases were detected.The re-sults showed that the method had good specificity with no cross reactivity with other pathogens exhibiting simi-lar symptoms.The minimum detection concentration was 1.55 copies/μL,and the sensitivity of this method was 10 times that of conventional PCR.The repeatability of the method was good with the coefficient of varia-tionsfor intra-and inter batch were from 0.79%to 1.83%.It indicated that the real-time fluorescent quantita-tive PCR method for ILTV detection established in this study had strong specificity and high sensitivity,which could provide an effective detection method for clinical diagnosis and epidemiological researches of ILTV.
Keywords：Chicken infectious laryngotracheitis virusTK geneReal-time fluorescent quantitative PCRSpecificitySensitivityRepeatability
Publication Date：2024-06-28
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：5( 128-132 )