不同PCR方法检测轮状病毒腹泻患儿血中RV基因  

Detection Rotavirus Genome in the Blood of Children with Rotavirus Diarrhea by Different Kinds of PCR Tests

作　　者：刘丽利[1] 黄永坤[2] 魏群德[2] 侯宗柳[3] 周丽芳[2] 王明珠[2] 

机构地区：[1]北京煤炭总医院儿科,北京100028 [2]昆明医学院第一附属医院儿科,昆明650032 [3]中国医学科学院医学生物学研究所,昆明650032

出　　处：《中国临床医学》2004年第3期427-429,共3页Chinese Journal of Clinical Medicine

摘　　要：目的 :探讨不同聚合酶链反应 (PCR)方法检测轮状病毒 (RV)腹泻患儿血中RVRNA敏感性以及病毒血症的情况。方法 :应用 2种逆转录聚合酶链反应 (RT -PCR)方法 ,对 30例轮状病毒腹泻患儿进行外周血血浆和单个核细胞中的RVRNA检测 ,阳性标本的PCR产物经琼脂糖电泳及测序证实。结果 :30例轮状病毒腹泻患儿 ,在外周血单个核细胞中检出RVRNA 1例 ,血浆中RVRNA检测均阴性 ,2种RT -PCR方法检测结果一致。阳性标本的PCR产物经测序结果证实属于人RVVP7片段序列 ,与G1型相似性为 94 % ,与G9型相似性为 75 %。检测出RVRNA阳性的病例有肠道外脏器损伤的表现。结论 :不同PCR方法可以检测到轮状病毒腹泻患儿血中RVRNA ,RV可能通过外周血单个核细胞向肠道外扩散 ,引起病毒血症造成肠道外脏器损伤。Objective: To investigate the sessitivity of RV RNA with different polymerase chair reaction tests and possibility of viremia in children with rotavirus diarrhea by measuring peripherial blood. Methods: The rotavirus RNA in the plasma samples and peripherial blood mononuclear cells (PBMCs) of 30 patients with rotavirus diarrhea were measured by two kinds of reverse transcriptase polymerase chain reaction (RT-PCR) tests. The positive PCR product was tested by agarose gel electrophoresis and was confirmed by analysing amino acid sequence. Results: One patient was found to be positive for RV RNA in the PBMCs and 30 patients were negative for RV RNA in the plasma. There were no difference between the results of two kinds of RT-PCR tests. The positive PCR product was confirmed as part of human RV VP7 by testing amino acid sequence. Identities of 94% with serotype G1 and 75% with G9 were noted on amino acid sequences. Extraintestinal symptoms were found in the positive case.Conclusion:RV RNA can be detected in the blood of children with rotavirus diarrhea by different kinds of PCR tests. Rotavirus may be transmitted by the PBMCs outside the intestinal tract and occur viremia which induces extraintestinal injury.

关 键 词：PCR 检测 轮状病毒 腹泻 小儿 血液 RV基因 病毒血症 聚合酶链反应 

分 类 号：R512.5[医药卫生—内科学]