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目的 建立检测甲肝患者外周血单个核细胞(PBMC)中甲肝病毒抗原 (HAV- Ag)的 APAAP方法 ,并对5 2例甲肝患者 PBMC中 HAV- Ag进行动态检测 .方法 应用抗 HAV- Mc Ab,羊抗鼠 Ig G及 APAAP建立了三层重叠的APAAP法 .对 5 2例甲肝患者急性期及恢复期 PBMC进行动态检测 ,对 HAV- Ag阳性率及平均阳性细胞百分率及其与甲肝病程的相关关系进行了分析 .结果 替代试验及阻断试验证明本方法特异性及敏感性好 ,结果可靠 .甲肝患者急性期PBMC中 HAV- Ag阳性率 (5 4.7% )及阳性细胞百分率[(14.2± 8.4) % ]明显高于恢复期 [6 .45 % ,P<0 .0 5 ;(1.8±0 .8) % ,P<0 .0 1];随着病程的延长 ,HAV- Ag阳性率及阳性细胞百分率逐渐下降 ,与病程存在显著负相关 (r=- 0 .99,P<0 .0 0 1;r=- 0 .98,P <0 .0 0 1) .结论 在甲肝病发过程中 ,患者 PBMC中有 HAV- Ag一过性出现 ,提示 HAV一过性侵犯了 PBMC.
Objective To establish an APAAP method for detection of hepatitis A virus antigen (HAV-Ag) in peripheral blood mononuclear cells (PBMCs) of hepatitis A patients and to detect HAV-Ag in PBMCs of 52 patients with hepatitis A. Methods Anti-HAV- Goat anti-mouse Ig G and APAAP established three-tier overlapping APAAP method.Animal and convalescent PBMC from 52 cases of hepatitis A were detected dynamically, and the correlation between the positive rate of HAV-Ag and the percentage of average positive cells and the course of hepatitis A The results of alternative tests and blocking tests proved that the specificity and sensitivity of this method is good and the results are reliable.The positive rate of HAV-Ag in PBMC of acute hepatitis A (5 4.7%) and the percentage of positive cells [(14.2 ± 8.4 )%] Was significantly higher than that of recovery [6 .45%, P <0.05; (1.8 ± 0.8)%, P <0.01] The percentage of positive cells decreased gradually and there was a significant negative correlation with the course of disease (r = - 0.99, P <0.01; r = - 0.98, P <0.001) In patients with PBMC HAV-Ag transient appeared, suggesting that HAV had a sexual abuse of PBMC.