论文部分内容阅读
目的：建立用地高辛（Ｄｉｇｏｘｉｇｅｎｉｎ，ＤＩＧ）标记的寡核苷酸基因探针鉴定Ａ群轮状病毒ＶＰ７Ｇ血清型的分子杂交技术，并应用该技术研究河南Ａ群轮状病毒的Ｇ血清型分布。方法：根据轮状病毒编码糖蛋白ＶＰ７的基因核苷酸序列，选择该基因高保守区序列中与ＶＰ７Ｇ血清型高度相关的核苷酸片段，人工合成，并用ＤＩＧ标记。在优选的杂交条件下进行杂交：结果：①Ｇ１、Ｇ２、Ｇ３和Ｇ４四种Ｇ血清型特异性寡核苷酸探针只与同血清型的轮状病毒参考毒株杂交，无交叉杂交现象。灵敏度和特异性达到放射性同位素３２Ｐ标记的水平。②１８８份经ＰＡＧＥ确定的轮状病毒阳性标本中，７９份（４２．０％）、３５份（１８．６％）、１５份（７．９％）分别与Ｇ１、Ｇ２或Ｇ３型探针杂交；未检出Ｇ４型；５６份（２９．８％）未能分型；３份分型特殊。结论：该Ｇ血清型分型技术具有高度特异性和灵敏度
OBJECTIVE: To establish a molecular hybridization technique for the identification of serogroup A of rotavirus A of group A using digoxigenin (DIG) labeled oligonucleotide probe and to study the G serotype distribution of group A rotavirus of Henan . Methods: According to the nucleotide sequence of rotavirus encoding glycoprotein VP7, we selected the highly conserved nucleotide sequence of VP7G and synthesized it with DIG. Hybridization was carried out under the optimal hybridization conditions. Results: ① The G-type G-type specific oligonucleotide probes of G1, G2, G3 and G4 hybridized only with the same serotype rotavirus reference strain without cross-hybridization. Sensitivity and specificity reach the level of radioisotope 32P labeling. ② Among the 188 positive samples confirmed by PAGE, 79 (42.0%), 35 (18.6%) and 15 (7.9%) were hybridized with G1, G2 or G3 probes, respectively ; G4 type was not detected; 56 (29.8%) failed to type; 3 special type. Conclusion: The G serotype typing technique has high specificity and sensitivity