应用免疫金银直接染色法诊断肾综合征出血热

A direct immunogold-silver staining method for diagnosing hemorrhagic fever with renal syndrome

应用免疫金银直接染色法（Ｄ－ＩＧＳＳ）对肾综合征出血热病毒（ＨＦＲＳＶ）感染的Ｖｅｒｏ－Ｅ６细胞进行ＨＦＲＳＶ抗原检测，发现Ｅ６细胞的胞浆和细胞膜上能清楚地观察到黑色均匀的银颗粒，而用ＨＦＲＳＶ免疫兔血清ＩｇＧ作阻断试验，却未见银颗粒。在此基础上对３２例ＨＦＲＳ患者检测外周血白细胞中的ＨＦＲＳＶ抗原，并和间接免疫荧光法（Ｉ－ＩＦＡ）进行对照，结果病程１０天以内Ｄ－ＩＧＳＳ法阳性率达到１０００％，个别病例病程２３天仍能检出ＨＦＲＳＶ抗原，而ＩＦＡ法在病程１１天以后阳性率明显下降，病程１４天后未检出ＨＦＲＳＶ抗原。说明Ｄ－ＩＧＳＳ法敏感性明显高于ＩＦＡ法。为对比外周静脉血和手指末稍血白细胞中ＨＦＲＳＶ抗原检出率，对３２例患者进行同步采血，结果ＨＦＲＳＶ抗原检出率无显著性差异（Ｐ＜００５）。我们认为Ｄ－ＩＧＳＳ法检测ＨＦＲＳ患者末稍血中的ＨＦＲＳＶ抗原，是简而易行的方法，可用于ＨＦＲＳ早期诊断。

A direct immunogold-silver staining (D-IGSS) method with 5nm colloidal gold-labeled rabbit anti-HFRS IgG antibodies was employed to detect HFRS virus antigen in the cytoplasm of white blood cells from HFRS patients and compared with indirect immunofluorescence assay (I-IFA). The results showed as follows: 1. The positive rate (87.5%) of HFRSV antigen detected by D-IGSS was significantly higher than that (65.6%) detected by I-IFA. in the white blood cells (P<0.05); 2. With the D-IGSS method the positive rates of HFRSV antigen in white blood cells of finger and vein blood were 81.3% and 87.5% respectively (P>0.05) with no significant difference but finger blood smears is more simple. We belive the D-IGSS mothod can be widely used for early diagnosis of HFRS at basic-level hospitals.