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目的建立检测版纳病毒(BAV)Ig G抗体的Alpha LISA方法。方法利用BAV重组VP9蛋白(GST-BAV VP9),以及制备的BAV兔抗血清和鼠抗血清,先将不同浓度的GST-BAV VP9和兔抗血清共同孵育,优化最佳抗原、抗体浓度;再用鼠抗血清与之竞争,建立竞争法检测方法,并应用于50份云南省发热病人BAV Ig G的检测。结果当GST-BAV VP9和兔抗血清浓度为1 nmol/L和1∶5 000时,结合力强并且可以被鼠抗血清竞争性抑制,即可用于BAV Ig G的检测。利用建立的方法,在50份发热病人血清中检测到9份阳性。结论建立了竞争性Alpha LISA方法,可用于BAV感染的筛查。
Objective To establish an Alpha LISA method for detection of BAV Ig G antibodies. Methods BAV recombinant VP9 protein (GST-BAV VP9), BAV rabbit antisera and mouse antisera were prepared. The optimal concentrations of GST-BAV VP9 and rabbit antisera were incubated with different concentrations of GST-BAV VP9 to optimize the optimal antigen and antibody concentration. Competition with mouse antiserum was established to establish competitive assay and to detect BAV Ig G in 50 febrile patients in Yunnan Province. Results When GST-BAV VP9 and rabbit antiserum concentration were 1 nmol / L and 1: 5000, the binding ability was strong and could be competitively inhibited by murine antiserum, which could be used for the detection of BAV Ig G. Using the established method, 9 positives were detected in sera from 50 febrile patients. Conclusion A competitive Alpha LISA method was established for the screening of BAV infection.