Chakraborty P, Barman N N, Sharma I
Department of Microbiology,Assam University,Silchar,Assam,India.
Department of Microbiology,College of Veterinary Science, Assam Agricultural University,Khanapara, Guwahati,Assam,India.
Epidemiol Infect. 2015 Sep;143(12):2503-11. doi: 10.1017/S0950268814003343. Epub 2015 Jan 9.
A restriction fragment length polymorphism (RFLP) assay was developed to examine the genetic relationship between 67 (29 Indian, 38 global) rotavirus isolates of human, bovine and porcine neonates. The assay involved direct digestion of RT-PCR amplified VP7 cDNAs with three restriction enzymes (VspI, HaeIII, NlaIV) independently. Forty-eight RFLP patterns were identified for all 67 strains, and of these 20 patterns were associated with Indian isolates. A correlation between the restriction patterns and G type was apparent through deduction of enzyme restriction sites from known sequences. Major G serotypes (G1, G2, G6, G8) with a few mixed types could be differentiated where there was a positive assortment of intrinsic serotypes from multiple host origin, and certain single or combined enzyme profiles were highly dominant in the population. Significant genetic variations were established between global and Indian isolates and none of the RFLP patterns were shared between them. These data suggest that the Indian wild-type rotavirus population is distinguishable based on the VP7 gene, and co-circulation of distinct strains in different hosts is foremost, indicating the possible likelihood of inter-species transmission.
开发了一种限制性片段长度多态性(RFLP)分析方法,以研究67株(29株来自印度,38株来自全球)人、牛和猪新生儿轮状病毒分离株之间的遗传关系。该分析方法包括分别用三种限制性内切酶(VspI、HaeIII、NlaIV)直接消化RT-PCR扩增的VP7 cDNA。在所有67株菌株中鉴定出48种RFLP模式,其中20种模式与印度分离株相关。通过从已知序列推导酶切位点,限制性模式与G型之间的相关性显而易见。在存在多种宿主来源的固有血清型的正向分类的情况下,可以区分主要的G血清型(G1、G2、G6、G8)以及少数混合类型,并且某些单一或组合的酶切图谱在群体中高度占主导地位。全球分离株和印度分离株之间存在显著的遗传差异,它们之间没有共享任何RFLP模式。这些数据表明,基于VP7基因,印度野生型轮状病毒群体是可区分的,并且不同宿主中不同菌株的共同循环是首要的,这表明种间传播的可能性很大。