2001—2003年杭州地区婴幼儿腹泻轮状病毒VP7型别分析

Analysis of VP7 serotype of rotavirus in infantile diarrhea in Hangzhou area from 2001 to 2003

目的研究杭州地区秋冬季婴幼儿腹泻轮状病毒(RV)的感染特点及RV分子流行病学特点。方法收集2001—2003年秋冬季来本院就诊拟诊为病毒性肠炎患儿的粪便共683份,采用乳胶凝集试验(LAT)检测A组RV,阳性者再用酶联免疫吸附试验(ELISA)分型检测RVVP7(G)血清型;将ELISA确定VP7(G)血清型的标本进一步做逆转录聚合酶链反应(RTPCR)扩增及RV基因测序,证实其VP7(G)血清型。结果683份标本中,RV阳性297份,检出率为43.5%(297/683);ELISA分型:VP7(G)血清型以G1、G3型为主,其中G1型占36.7%(109/297)、G3型占30.9%(92/297),其余G2、G4、混合型和未分型分别占4.0%(12/297)、7.1%(21/297)、11.8%(35/297)和9.4%(28/297);每年流行的VP7(G)血清型的主流不同,2001年以G1型为主,占54.9%(45/82)、G3型占14.6%(12/82),2003年以G3型为主,占43.2%(63/146)、G1型占29.5%(43/146)。RTPCR扩增、基因序列测定和同源性分析证实了ELISA分型的可信性。结论2001—2003年杭州地区秋冬季婴幼儿腹泻RVVP7(G)血清型以G1、G3型为主,且每年流行的主流血清型不同。

Objective To investigate molecular epidemiologic features of rotavirus （RV） infection in infantile diarrhea in Hangzhou area. Methods Stool specimens of 683 infants with suspected acute viral enteritis in the autumn and winter of 2001--2003 were collected. RV（ group A）was detected by using latex agglutination test （LAT）. VP7 serotype （G） positive specimens were detected by using enzyme linked immunosorbent assay （ ELISA ） and then the RNA of the virus was determined with reverse transcription polymerase chain reaction （ RT-PCR ）. cDNA of VP7 gene fragment was sequenced by automatic gene analyzor （ABI3730） and compared with the RV VP7 gene sequences stored in Genebank. Results RV was detected in 297 of 683（43.5% ）specimens by LAT. The highest frequency of RV （group A） detected was 52.9% （228/431） in patients aged 7--18 months. The prevalent serotypes were G1 （36. 7% , 109/297 ）and G3 （ 30. 9% , 92/297 ） , followed by mixed type （ 11.8% , 35/297 ） , untyped （ 9. 4% , 28/297 ） , G4（7. 1% ,21/297） and G2（4.0%, 12/297）. The prevalent serotypes seen each year were different. G1（54.9% , 45/82） was the major serotype in 2001 followed by G3 （ 14. 6% , 12/82）. In 2003, the major serotype was G3（43.0% ,63/146） and followed by G1 （29.5% ,43/146）. The reliability of ELISA was confirmed by RT-PCR, gene sequencing and homology analysis. Conclusion The main prevalent serotypes of VP7 of rotavirus were G1 and G3. The dominant serotypes of rotavirus varied in Hangzhou area from 2001 to 2003.