Kwon H M, Jackwood M W, Gelb J
Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602.
Avian Dis. 1993 Jan-Mar;37(1):194-202.
Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis were used to differentiate between serotypes of several infectious bronchitis virus (IBV) strains. A sequence of 1720 base pairs (bp) that contains the S1 glycoprotein gene of IBV was amplified by PCR, purified, and digested with restriction enzymes. Eleven reference IBV strains were grouped according to the RFLP patterns. The IBV Holte, Arkansas DPI, SE 17, Md 27, and Iowa 97 strains could be differentiated from the other IBV strains using the restriction enzyme HaeIII. The Beaudette, Massachusetts 41, Connecticut, and Florida 88 strains had the same HaeIII RFLP pattern but could be differentiated using XcmI and BstYI restriction enzymes. The Gray and JMK strains could not be differentiated by their RFLP patterns following digestion with 23 different restriction enzymes. Twenty-six samples (field isolates and reference strains) of IBV, previously serotypes by the virus-neutralization (VN) test in embryonating eggs, were analyzed in a blind fashion. The results using the PCR and RFLP analysis agreed with the serotype for traditional and variant IBV viruses as determined by the VN test.
采用聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析来区分几种传染性支气管炎病毒（IBV）毒株的血清型。通过PCR扩增出一段包含IBV S1糖蛋白基因的1720个碱基对（bp）的序列，进行纯化后用限制性内切酶消化。根据RFLP模式对11株参考IBV毒株进行分组。使用限制性内切酶HaeIII可将IBV Holte、阿肯色州DPI、SE 17、Md 27和爱荷华97毒株与其他IBV毒株区分开来。Beaudette、马萨诸塞州41、康涅狄格和佛罗里达88毒株具有相同的HaeIII RFLP模式，但可使用XcmI和BstYI限制性内切酶进行区分。在用23种不同的限制性内切酶消化后，Gray和JMK毒株无法通过其RFLP模式进行区分。对26份IBV样本（田间分离株和参考毒株）进行了盲法分析，这些样本之前在鸡胚中通过病毒中和（VN）试验确定了血清型。PCR和RFLP分析的结果与VN试验确定的传统和变异IBV病毒的血清型一致。
