Objective Chemiluminiscence is used to detect the reactivity of 4 subtypes prokaryotic recombinant HIVI gp41 extracellular proteins to pos. and neg. samples.The results could indirectly reflect the antigen epitope exposure status and provide a method to analyze the abnormal samples on scientific and clin. research.Methods A total of 4 gp41 gene fragments of HIVI domestic mainstream genotypes( BC,AE,B,C) were selected and constructed to PGEX4 T-3 vector by PCR and restriction enzyme digestion.The optimized purification method was used to obtain HIVI extramembrane gp41 proteins which worked as coating antigens.Then,the coating antigens and com. gp41-HRP were used in the sandwich ELISA to estimate the reactivity of HIVI neg. and pos. samples.Thereby the exposure statuses of the corresponding protein epitopes were judged.Results The gp41 extramembrane proteins which were selected from the same extramembrane region in different genotypes could express target protein with expected mass weight via p GEX4 T-3 prokaryotic vector.The purified proteins work as coating protein with the same concentration,proceed to test HIVI neg. and pos. samples,to evaluate the epitope exposure status of gp41 protein which was expressed by prokaryotic system.Conclusion The prokaryotic expression system efficiently expressed the gp41 outer membrane protein of HIV type I mainstream subtype.The chemiluminescence method was used to construct the sandwich method to test neg. pos. samples.The reactivity was inconsistent,which indirectly reflected that different genotypes selected the same expression region for recombinant expression in vitro.There are differences in structural folding methods,and this study provides the basis for HIV research and clin. specimen verification.