脊灰病毒分离和鉴定新检测流程的应用

Application of poliovirus isolation and identify new measurement procedures

目的缩短检测脊灰病毒的时限,及时发现河北省可能出现的脊灰疫苗衍生病例(VDPV)。方法对河北省2013年急性弛缓性麻痹(AFP)病例及其密切接触者的粪便标本用L20B和RD两种细胞进行病毒分离,用实时荧光定量逆转录-聚合酶链反应(Real Time Fluorescent Quantitative Reverse Transcription-polymerase Chain Reaction,rRTPCR)对分离到的L20B阳性分离物进行型内鉴定和血清型鉴定。结果从送检的334例AFP病例及接触者粪便标本中检测出L20B阳性分离物26株,用real-time PCR法进行了病毒血清型鉴定和型内鉴定,共监测到脊灰疫苗类似株(sabin-like,SL)19株,其中Ⅰ型9株、Ⅱ型1株、Ⅲ型6株、混合型3株;将混合型进行单型分型后共得24株,进行VP1区核苷酸序列测定,发生高变异的两株出现在Ⅰ型,均为6个核苷酸变异。结论 2013年分离到的脊灰疫苗病毒有部分变异,没有发现疫苗衍生脊灰病毒。

Objective Shortening the time limit of test of poliovirus,to detect vaccine derived polio viruses（ VDPVs） timely in Hebei province. Methods The polioviruses were isolated from stool samples of acute flaccid paralysis（ AFP） cases from 2013 by using L20 B and RD cell,real time fluorescent quantitative reverse transcription- polymerase chain reaction（ rRT- PCR） was applied in intratypic differentiation. Results Detected in stool samples from 334 cases of AFP cases and contacts censorship of the L20B- positive isolates 26.Virus serotypes were identified and the type identified by real- time PCR method of monitoring the polio vaccine strains similar（ sabin- like,SL） 19 strains,among which type Ⅰ19,typeⅡ1,type Ⅲ 6,and mixed type 3; put mixed type to monotype partying got 24 in total,after VP1 region nucleotide sequence determination,the occurrence of 2 high variability in typeⅠ,are 6 nucleotide variations. Conclusions Polio vaccine virus isolated in 2013 had some of the variation,found no vaccine- derived poliovirus.