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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 To shorten the time limit for detecting poliovirus and find out the possible cases of poliovirus vaccine (VDPV) in Hebei Province. Methods Stool samples of acute flaccid paralysis (AFP) cases and their close contacts in Hebei Province in 2013 were isolated by using L20B and RD cells. Real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction (Real Time Fluorescent Quantitative Reverse Transcription-polymerase Chain Reaction, rRTPCR) were used to identify and serotype the isolated L20B positive isolates. Results Totally 26 L20B positive isolates were detected from 334 cases of AFP and contact stool samples. The virus serotypes and in-cell types were identified by real-time PCR. 19 strains were type Ⅰ, 1 were type Ⅱ, 6 were type Ⅲ and 3 were mixed type. A total of 24 strains were obtained after single type typing, and VP1 nucleotide Sequence analysis showed that two strains with high variation appeared in type Ⅰ, which were all six nucleotide variations. Conclusions There was a partial variation of the poliovirus isolated in 2013 and no vaccine-derived poliovirus was found.