Razafindratsimandresy R, Rakoto Andrianarivelo M, Rousset D
Institut Pasteur de Madagascar, BP 1274, 101 Antananarivo, Madagascar.
Arch Inst Pasteur Madagascar. 2003;69(1-2):27-32.
The differentiation of the vaccine or wild origin of Poliovirus at the laboratory is an important step towards the process of the poliomyelitis eradication. We report herein the results obtained from Poliovirus types 3 and 2, isolated in Madagascar in 1997 and 2002 from healthy children and cases of acute flaccid paralysis, respectively. The technique used is based on the amplification of genome (RT-PCR), followed by Restriction Fragment Length Polymorphism assay (RFLP), performed in 3 different regions of the genome. In the capsid region (VP3-VP1 and VP1-2A), RFLP analysis allowed us to differentiate without ambiguity the wild or vaccine origin of the Poliovirus type 3, and to identify Vaccine-Derived Poliovirus (VDPV) type 2. In the noncapsid region, including the RNA polymerase and 3' non coding region (3Dpol-3' NTR), the VDPV were found to be recombinant with other Enteroviruses. These results confirm that RFLP assay is a reliable tool for intratypic differentiation and to study the genetic drift and recombination of Poliovirus.
在实验室鉴别脊髓灰质炎病毒的疫苗株或野生株来源是迈向根除脊髓灰质炎进程的重要一步。我们在此报告1997年和2002年分别从马达加斯加健康儿童和急性弛缓性麻痹病例中分离出的3型和2型脊髓灰质炎病毒的检测结果。所采用的技术基于基因组扩增(逆转录聚合酶链反应),随后在基因组的3个不同区域进行限制性片段长度多态性分析(RFLP)。在衣壳区域(VP3 - VP1和VP1 - 2A),RFLP分析使我们能够明确区分3型脊髓灰质炎病毒的野生株或疫苗株来源,并鉴定出2型疫苗衍生脊髓灰质炎病毒(VDPV)。在非衣壳区域,包括RNA聚合酶和3'非编码区(3Dpol - 3'NTR),发现VDPV与其他肠道病毒发生了重组。这些结果证实,RFLP分析是进行型内鉴别以及研究脊髓灰质炎病毒基因漂移和重组的可靠工具。