以合成肽为抗原检测人巨细胞病毒抗体的酶联免疫吸附试验的建立及评价

Establishment and evaluation of an indirect enzyme-linked immunosorbent assay based on synthetic peptides for the detection of specific antibodies to Human cytomegalovirus

目的以合成多肽为抗原建立用于人巨细胞病毒(HCMV)抗体检测的间接酶联免疫吸附试验(ELISA)。方法以固相多肽合成技术合成了HCMV 7个抗原性肽段,并对其抗原性进行了初步评价,选择高抗体反应的肽段为抗原,建立检测HCMV特异性抗体的间接ELISA。结果7个肽段中,抗体反应性较高肽段有4个;以合成多肽为抗原的间接ELISA对HCMV IgG抗体检测的敏感性和特异性分别为90.9%和100.0%,对于IgM抗体检测的敏感性和特异性分为90.0%和100.0%;与国外试剂盒测定结果有良好的符合性;该方法的精密度及稳定性良好。结论合成的HCMV多肽片段可作为抗原用于HCMV的检测,以此多肽为抗原建立的间接ELISA可用于临床HCMV特异性抗体的检测。

Objective To establish an indirect enzyme-linked immunosorbent assay （ELISA） for the detection of Human cytomegalovirus （HCMV） specific antibodies, based on synthetic antigenic peptides of HCMV. Methods 7 highly antigenic peptides were synthesized by solid phase peptide synthesis technique, and their antibody reactivity was evaluated by indirect ELISA. Some peptides which had highly antibody reactivity were selected as the antigens and an indirect ELISA was established for the detection of specific antibodies to HCMV. Results There were 4 peptides which had high antibody reactivity among those 7 peptides ; The sensitivities of the indirect ELISA based on the combination of synthetic peptides were 90.9% and 90.0% for the detection of HCMV IgG antibodies and IgM antibodies, and the specificities were both 100.0%. Compared with imported HCMV test kits, there was a good accordance between two assays. The assay had good precision and stability. Conclusions The synthetic peptides could be used as antigens for the detection of HCMV specific antibodies and the indirect ELISA based on these peptides could be used in the detection of HCMV specific antibodies in clinical diagnosis of HCMV infection.