Chen H Y, Zhou Q, Zhang M F, Giambrone J J
Department of Biochemistry and Molecular Biology, College of Biological Sciences, China Agricultural University, Beijing, P. R. China.
Avian Dis. 1998 Oct-Dec;42(4):762-9.
The VP2 hypervariable region of infectious bursal disease virus (IBDV) from nine Mainland Chinese strains was amplified by reverse transcriptase/nested polymerase chain reaction and cloned into pGEM-T vector. The nine isolates, which were from the center (HN3), the north (Bj-1, B2/28, HD96), the east (JS-18 and AH-2), the northeast (D11-2, C4-2), and the west (Ts) of China, were sequenced and compared with each other and with six reference IBDV sequences. Clustering analysis separated the nine isolate into two groups. The six virulent isolates, propagated in bursae, formed the first group. They revealed only one to three amino acid changes from the very virulent (vv) European and Japanese isolates, suggesting that they might have the same origin as European and Japanese vvIBDV strains. On the basis of their distinct geographic origins, extensive dissemination of vvIBDV in China was indicated. (The other three chicken embryo fibroblast cell cultured isolates with mild pathogenicity were placed in the second group.) Their sequences correlated closely with those of the culture-adapted strains (Cu-1 (4) and Cj-801). None of the nine isolates showed very close sequence relationship with the antigenic variant strains from the USA. Although antigenic variants have been reported in China, the reverse transcriptase/polymerase chain reaction-restriction endonuclease analyses of the nine viruses tested herein were not similar to any U.S.A. variant strains on the basis of computer software analysis. Our results and conclusions agree with a previous molecular study of IBDV isolates from the south of China.
通过逆转录/巢式聚合酶链反应扩增了来自中国大陆9个毒株的传染性法氏囊病病毒(IBDV)的VP2高变区,并将其克隆到pGEM-T载体中。这9个分离株分别来自中国的中部(HN3)、北部(Bj-1、B2/28、HD96)、东部(JS-18和AH-2)、东北部(D11-2、C4-2)和西部(Ts),对它们进行了测序,并相互比较,同时与6个IBDV参考序列进行了比较。聚类分析将这9个分离株分为两组。在法氏囊中增殖的6个强毒株形成了第一组。它们与超强毒(vv)欧洲和日本分离株相比,仅显示出1至3个氨基酸的变化,这表明它们可能与欧洲和日本的vvIBDV毒株有相同的起源。基于它们不同的地理来源,表明vvIBDV在中国广泛传播。(另外3个在鸡胚成纤维细胞中培养的致病性温和的分离株被归入第二组。)它们的序列与适应细胞培养的毒株(Cu-1(4)和Cj-801)密切相关。这9个分离株中没有一个与来自美国的抗原变异株显示出非常密切的序列关系。尽管中国已报道有抗原变异株,但根据计算机软件分析,本文检测的9种病毒的逆转录/聚合酶链反应-限制性内切酶分析结果与任何美国变异株均不相似。我们 的结果和结论与先前对来自中国南方的IBDV分离株的分子研究一致。