Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model.
Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model.
复制标题
DOI：
10.1007/s00705-009-0357-1
复制
发表时间：
2009
影响因子：
2.7
通讯作者：
Ignjatovic J
中科院分区：
文献类型：
--
作者：
Hewson K;Noormohammadi AH;Devlin JM;Mardani K;Ignjatovic J
传染性支气管炎病毒（IBV）是一种冠状病毒，可引起家禽的上呼吸道、肾脏和/或生殖系统疾病，发病率很高。IBV的分类对于实施预防接种策略以控制商业家禽中的疾病是重要的。目前，IBV S1基因序列分析的冗长过程被认为是IBV毒株鉴定的金标准，高核苷酸同一性（例如≥95%）表明相关毒株。然而，该基因具有突变和/或经历重组的高倾向，并且单独地其对于菌株鉴定可能不可靠。根据鸡传染性支气管炎病毒（IBV）3′非翻译区（UTR）序列，建立了实时荧光定量PCR（RT-PCR）结合高分辨熔解曲线（HRM）分析的方法，用于商业家禽中IBV的快速检测和分类。HRM曲线产生的230至435-bp的几个IBV株的PCR产物进行了进一步的分析，使用的数学模型也在本研究中开发。结果表明，HRM曲线分析与数学模型相结合，可将190对IBV毒株中的189对按3′UTR和S1基因同源性进行可靠分组。新建立的RT-PCR/HRM曲线分析模型能够快速检测和鉴定IBV新毒株和疫苗相关毒株。该模型是一种快速、可靠、准确和非主观的检测鸡传染性支气管炎病毒的系统。
Infectious bronchitis virus (IBV) is a coronavirus that causes upper respiratory, renal and/or reproductive diseases with high morbidity in poultry. Classification of IBV is important for implementation of vaccination strategies to control the disease in commercial poultry. Currently, the lengthy process of sequence analysis of the IBV S1 gene is considered the gold standard for IBV strain identification, with a high nucleotide identity (e.g. ≥95%) indicating related strains. However, this gene has a high propensity to mutate and/or undergo recombination, and alone it may not be reliable for strain identification. A real-time polymerase chain reaction (RT-PCR) combined with high-resolution melt (HRM) curve analysis was developed based on the 3′UTR of IBV for rapid detection and classification of IBV from commercial poultry. HRM curves generated from 230 to 435-bp PCR products of several IBV strains were subjected to further analysis using a mathematical model also developed during this study. It was shown that a combination of HRM curve analysis and the mathematical model could reliably group 189 out of 190 comparisons of pairs of IBV strains in accordance with their 3′UTR and S1 gene identities. The newly developed RT-PCR/HRM curve analysis model could detect and rapidly identify novel and vaccine-related IBV strains, as confirmed by S1 gene and 3′UTR nucleotide sequences. This model is a rapid, reliable, accurate and non-subjective system for detection of IBVs in poultry flocks.
登录
查看更多内容
影响因子：
5
作者：
Cavanagh D;Davis PJ;Mockett AP
通讯作者：
Mockett AP
高分辨率熔化用于TP53外显子5-8的突变扫描。
影响因子：
3.8
作者：
Krypuy, Michael;Ahmed, Ahmed Ashour;Etemadmoghadam, Dariush;Hyland, Sarah J.;deFazio, Anna;Fox, Stephen B.;Brenton, James D.;Bowtell, David D.;Dobrovic, Alexander
通讯作者：
Dobrovic, Alexander
影响因子：
14.9
作者：
Benson, Dennis A.;Karsch-Mizrachi, Ilene;Sayers, Eric W.
通讯作者：
Sayers, Eric W.
影响因子：
2.8
作者：
Liu, Shengwang;Han, Zongxi;Rong, Jungong
通讯作者：
Rong, Jungong
影响因子：
5
作者：
WILLIAMS, AK;WANG, L;COLLISSON, EW
通讯作者：
COLLISSON, EW