Identification of VP7 Serotypes of Human Rotaviruses by Enzyme-Linked Immunosorbent Assay and Reverse Transcription-Polymerase Chain Reaction

以酶連免疫試驗及反轉錄－聚合酶連鎖反應鑑定人類輪狀病毒VP7血清型

李君男(Chun-Nan Lee) ； 高全良(Chuan-Liang Kao) ； 甯孝真(Hsiao-Chen Ning) ； 傅學琳(Hsieh-Lin Fuh) ； 李慶雲(Chin-Yun Lee)

《Acta Paediatrica Sinica》 38卷6期 (1997/12) Pp. 454-462

https://doi.org/10.7097/APS.199712.0454

摘要

台灣地區之人類輪狀病毒屬於何種血清型仍不甚清楚。本研究建立了三種鑑定VP7(G)血清型之方法，即螢光點中和試驗(FFN)、以G血清特異之單株抗體進行酶連免疫試驗(ELISA-MAb)、與反轉錄－聚合酶連鎖反應(RT-PCR)。以FFN爲鑑定G血清型之標准參攷方法，當以ELISA-MAb和RT-PCR來鑑定輪狀病毒分離株之G血清型時，顯示結果正確而且敏感度較FFN爲高。然而當直接應用於臨床檢體時，ELISA-MSb則不甚理想，在264個檢體中僅有65.5%可定出血清型。而由RT-PCR預測G血清型之度則可碠89.4%。如將兩種方法合併計算則93.2%之檢體可定出G血清型。鑑定G血清型的結果可與RNS電泳圖譜相互對應，如輪狀病毒具有相似的RNA電泳型則其血清型均相同，故RNA電泳漙可做爲參攷發預測部分檢體輪狀病毒之G血清型。因酶連免疫試驗之操作簡便且成本較低，適於做爲篩檢之方法。至於以每連銀川市試驗無法定出G血清型這檢體則可利用RT-PCR的方法。此外，對於一些末擁有G血清型特異之單株抗體的實驗室，則RT-PCR特別有利用之價值。

關鍵字

輪狀病毒 ； VP7血清型 ； 酶連免疫試驗 ； 反轉錄－聚合酶連鎖反應

並列摘要

Serotype specificity of human rotaviruses in Taiwan remained unclear. This study has established three methods for identification of G serotypes, fluorescent focus neutralization test (FFN), enzyme- linked immunosorbent assay by incorporating VP7 (G) serotype-specific monoclonal antibodies (ELISAMAb) and reverse transcription-polymerase chain reaction (RT-PCR). FFN is a reference standard test for determination of G serotype. RT-PCR and ELISA-MAb were used for serotyping of rotavirus isolates and appeared to be accurate and more sensitive than FFN. Nevertheless, for serotyping. the rotaviruses present in clinical stool samples, ELISA-MAb was not satisfactory, only 65.5% of the 264 specimens could be serotyped. The sensitivity of serotype prediction by RT-PCR was 89.4%. When both ELISAMAb and RT-PCR were taken into account, G serotype could be predicted for 93.2% of the specimens. Serotyping results were comparable with the RNA profiles. Rotaviruses with similar RNA electrophoretic pattern had the same serotype specificity. RNA patterns could be used as a reference for prediction of serotypes. Because ELISA is easy to perform and less expensive, it would be suggested as a screening test. RT-PCR could be used for samples which could not be serotyped by ELISA-MAb. Furthermore, RT-PCR is especially valuable when serotype-specific monoclonal antibodies are not available.

並列關鍵字

rotavirus ； VP7serotype ； ELISA ； RT-FCR