免疫金银染色检测双股DNA抗体的研究

THE STUDY ON DETECTION OF ANTI-dsDNA ANTIBODY BY IMMUNOGOLD-SILVER STAINING TECHNIQUE

首次报告以酸洗脱小鼠肝、肾印片(AES-肝、肾)及酶处理人精子头核(ESN)为双股DNA底物、以免疫金银染色(IGSS)检测双股DNA抗体(AdsDNA)。与免疫荧光试验(IFT)的平行对照表明,无论用何种底物,IGSS的敏感性均显著高于IFT,而特异性与IFT相同。AES-肝、AES-肾及ESN三底物间的平行对照表明,其AdsDNA检出结果符合率无论在IGSS或IFT法均>80%,其特异性基本相近,敏感性以AES-肾和ESN较高。此三种底物与目前多用的短膜虫(CL)的平行对照表明,其检出符合率均>84％,特异性与敏感性均与CL相似。作者认为AES-肝/肾或ESN为底物的IGSS检测AdsDNA技术简便实用、特异敏感,值得推广。

A new method was developed for detection of the anti-dsDNA antibody (AdsDNA)by using immunogold-silver staining technique ( IGSS ) and various antigen substrates, i. e. HCl-treated mouse liver/kidney imprints ( AES-liver /kidney ) and chymotrypsin and trypsin-treated human spermatozoa ( ESN ) . This method was compared with the current ones in respect to the method for staining and the selection of dsDNA antigen substrate. The results showed that compared with immunofluorescence test ( IFT ) , IGSS has significantly high sensitivity ( p<0.05 ) and similar specificity ( p>0.05 ). Study of the 3 substrates (using IGSS and IFT ) showed they were similar in specificity but AES-liver was slightly less sensitive than AES-kidney and ESN (P<0.05). Compared with the popular antigen substrate crithidia luciliae ( CL ) , the 3 substrates could give similar positive rate ( >84% ) and comparable specificity and sensitivity. Since the new method is equally specific and sensitive, but costs less and is easier to carry out than the current ones, it should be more practicable and worth wide adaptation.