摘要
目的探讨单管巢式聚合酶链反应（ＳＮＰＣＲ）检测石蜡包埋组织结核分支杆菌ＤＮＡ的特异性和敏感性。方法应用普通ＰＣＲ（ＧＰＣＲ）、双管巢式ＰＣＲ（ＤＮＰＣＲ）和ＳＮＰＣＲ对结核分支杆菌ＢＣＧ和３０例结核性淋巴结炎石蜡包埋组织进行结核分支杆菌复合群ＩＳ６１１０特异插入序列片段ＤＮＡ检测。结果ＤＮＰＣＲ和ＳＮＰＣＲ检测ＢＣＧＤＮＡ均于１５ｆｇ以上呈现阳性结果，其敏感性明显优于ＧＰＣＲ（４８０ｆｇ）。ＧＰＣＲ、ＤＮＰＣＲ和ＳＮＰＣＲ检测３０例结核性淋巴结炎阳性率分别为４３％、１００％和１００％，抗酸染色阳性率（１０％）与３种ＰＣＲ法相比差异有非常显著意义（均Ｐ＜０．０１）。ＧＰＣＲ阳性率与ＤＮＰＣＲ和ＳＮＰＣＲ相比差异亦具非常显著意义（均Ｐ＜０．０１）。ＳＮＰＣＲ阳性率与ＤＮＰＣＲ相同。结论巢式ＰＣＲ检测淋巴结石蜡包埋组织结核分支杆菌的敏感性显著高于ＧＰＣＲ，其中ＳＮＰＣＲ具有与ＤＮＰＣＲ相同的特异性和敏感性，并具有更大的实用价值。
Objective To study the sensitivity and specificity of single tube nested polymerase chain reaction (SN PCR) technique in detecting Mycobacterium tuberculosis DNA in paraffin embedded tissues. Method BCG DNA and the paraffin embedded tissues of 30 cases with typical tuberculous lymphadenitis were examined to detect the IS6110 specific insertion sequence DNA of Mycobacterium tuberculosis complex by general PCR(G PCR), double tube nested PCR (DN PCR) and SN PCR techniques. Result Fifteen fg or more BCG DNA could show positive results by DN PCR and SN PCR techniques, while 480 fg by G PCR method. The positive rates of G PCR, DN PCR, and SN PCR in detecting Mycobacterium tuberculosis DNA in 30 cases with tuberculous lymphadenitis were 43%, 100%, and 100% respectively, all of which were significantly different ( P <0.01) from that of the acid fast stainning method (10%). Significant differences in the positive rates also existed between G PCR and DN PCR, G PCR and SN PCR (both P <0.01), while the positive rates of DN PCR and SN PCR were found same. Conclusion The sensitivities of nested PCR techniques in detecting Mycobacterium tuberculosis are significantly higher than that of G PCR, whereas the sensitivities as well as the specificities of SN PCR and DN PCR are the same. Thus SN PCR seems more practicable.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
1998年第7期399-401,共3页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
天津市自然科学基金
关键词
淋巴结结核
结核杆菌
聚合酶链反应
DNA
Mycobacterium tuberculosis Polymerase chain reaction Tuberculosis, lymph node
