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目的:探讨改良免疫组化染色法在诊断乙型肝炎病毒相关性肾炎(HBV-GN)的应用前景。方法:改良免疫组化染色法采用高温高压+XXⅣ型蛋白酶消化的抗原修复法,行一抗和二抗两步孵育法,其中二抗与酶标多聚体相连。病人肾组织标本同时行免疫荧光染色法检测作对照。结果:(1)40例血清学HBsAg阳性患者的肾穿组织标本中,改良免疫组化染色法HBsAg阳性27例,阳性率为67.5%;HBcAg阳性7例(其中HbsAg同时阳性6例,HbsAg阴性1例),阳性率为17.5%;HbsAg、HBcAg同时阴性12例。免疫荧光染色法HBsAg阳性29例(72.5%);HBcAg阳性10例(25.0%,均同时呈HbsAg阳性);HbsAg、HBcAg同时阴性11例。(2)两种染色比较,HbsAg阳性率相近(改良组化67.5%vs荧光72.5%),其中同时阳性的有22例,同时阴性的5例,总一致率为67.5%(27/40),阳性一致率在改良免疫组化染色中为81.5%(22/27),在免疫荧光染色中为75.9%(22/29)。(3)10例血清丙氨酸转氨酶升高的患者中,改良免疫组化HbsAg染色阳性9例,免疫荧光染色阳性7例;血白蛋白<30g/L而尿Pro定量未达到>3.5g/24h的患者7例;6例病理呈膜性或/和膜增殖性肾炎(其中改良组化染色阳性5例,荧光染色阳性3例)。有2例行肝组织穿刺活检,病理显示肝脏病变。结论:改良免疫组化染色可提高HBV-GN的乙肝抗原检测率,有临床应用推广价值。
Objective: To explore the application of modified immunohistochemical staining in the diagnosis of hepatitis B virus associated nephritis (HBV-GN). Methods: Immunohistochemical staining was performed by the method of high temperature and high pressure + XXIV protease digestion, followed by two-step incubation of primary antibody and secondary antibody, in which secondary antibody was linked to enzyme-labeled polymer. Patients with renal tissue samples simultaneously detected by immunofluorescence staining as a control. Results: (1) HBsAg positive was found in 40 cases of renal biopsy specimens from HBsAg-positive patients (n = 27) with modified immunohistochemical staining, the positive rate was 67.5%; HBcAg positive was 7 (HbsAg positive 6, HbsAg negative 1 case), the positive rate was 17.5%; HbsAg, HBcAg negative at the same time in 12 cases. Immunofluorescence staining of HBsAg positive in 29 cases (72.5%); HBcAg positive in 10 cases (25.0%, both positive HbsAg); HbsAg, HBcAg negative at the same time in 11 cases. (2) Compared with the two kinds of staining, the positive rate of HbsAg was 67.5% vs 72.5%, of which 22 cases were positive at the same time and 5 cases were negative at the same time. The total coincidence rate was 67.5% (27/40) The positive agreement rate was 81.5% (22/27) in modified immunohistochemical staining and 75.9% (22/29) in immunofluorescence staining. (3) Of the 10 patients with elevated serum alanine aminotransferase, 9 were positive for HbsAg staining by modified immunohistochemistry and 7 were positive by immunofluorescence staining; those with serum albumin <30 g / L and urine Pro were less than 3.5 g / 7 cases in 24h; 6 cases were pathological membranous or / and proliferative glomerulonephritis (including modified positive staining in 5 cases, positive staining in 3 cases). There are 2 cases of liver biopsy, the pathology showed liver disease. Conclusion: Improved immunohistochemical staining can improve the detection rate of HBV-GN hepatitis B antigen and has clinical application value.