恙虫病东方体三种膜蛋白抗原包被酶联免疫吸附试验的特异性和敏感性分析

ANALYSES OF SPECIFICITY AND SENSITIVITY OF ENZYME-LINKED IMMUNOSORBENT ASSAY WITH THREE MEMBRANE-ASSOCIATED PROTEINS OF ORIENTIA TSUTSUGAMUSHI

用恙虫病东方体的r47、 r56、 r58 kDa重组膜蛋白抗原包被建立酶联免疫吸附试验（ ELISA ），对恙虫病东方体实验感染小鼠血清和临床患者血清做IgG抗体检测。8份恙虫病感染小鼠血清中除1份血清与r58 kDa蛋白的反应阴性外其他反应均为阳性；而这3个重组蛋白与贝氏柯克斯体（ Q热病原体）、莫氏立克次体（地方性斑疹伤寒病原体）、黑龙江立克次体（远东斑点热病原体）感染小鼠血清反应均为阴性。另外，12份恙虫病患者血清对这3个蛋白的反应均有11份（92％）为阳性。另外， r47 kDa抗原对11份Q热患者血清、11份斑疹伤寒患者血清、13份斑点热患者血清的反应阳性率分别有27％、18％、23％，对r56 kDa抗原分别为27％、0％、8％，对r58 kDa抗原分别为18％、18％、23％。这些结果显示这3个重组蛋白抗原均有识别恙虫病东方体感染血清的良好特异性和敏感性，用这3种重组蛋白抗原建立的ELISA将为我国恙虫病血清学诊断提供新的实用方法。

The enzyme-linked immunosorbent assay （ ELISA） with a recombinant outer membrane protein （ r47, r56, or r58 kDa） of Orientia tsutsugamushi was analyzed for specificity and sensitivity in detection of specific Ig G antibodies in sera from mice and patients suffered from O.tsutsugamushi infection.As a result, all of 8 serum specimens from mice experimentally infected with O.tsutsugamushi positively reacted with r47, 56 and r58 kDa protein respectively, except for that one serum negatively reacted with r58 kDa, while all of 8 sera from mice infected with Coxiella burnetii, Rickettsia mooseri or R.heilongjiangensis negatively reacted with the three proteins.In addition, 11 of 12 sera from patients with tsutsugamushi disease positively reacted with the three proteins, respectively, while 11 sera from patients with Q fever, 11 sera from patients with endemic typhus, and 13 sera from patients with spotted fever positively reacted to r47 kDa in the ratios of 28%, 18%, and 23%, to r56 kDa in the ratios of 28%, 0%, and 8%, and to r58 kDa in the ratios of 18%, 18%, and 23%, respectively.The results suggest that the ELISA coated with r47, r56, or r58 kDa has a good specificity and sensitivity for serological diagnosis of tsutsugamushi disease.