Crisóstomo Vázquez P, Jiménez Cardoso E
Laboratorio de Investigación en Parasitología, Hospital Infantil de México, Federico Gómez.
Rev Invest Clin. 2000 May-Jun;52(3):255-60.
Cysteine-proteinase of Entamoeba histolytica have been considered implicated like important virulence factors in the pathogenesis of amebiasis. On the basis of the differences in ethnic gene that encodes to 30 kDa proteinase. The present study validated a strategy to differentiate strains of pathogenic and non-pathogenic Entamoeba histolytica by restriction patterns.
Thirteen stool samples with Entamoeba histolytica cyst from 4 asymptomatic and 9 symptomatic patients ages and sex different into Robinson' medium were used. DNA obtained was used by amplified gene ethnic and it was cut with restriction enzyme Taq I and Hinf I.
All strains were cultivated into Robinson's medium. A 530 bp fragment which hybridated with probe for Entamoeba histolytica was obtained. By the way valuation by restriction patterns with Taq I and Hinf I show that two of four samples of asymptomatic patients belong to pathogenic strain. It agrees with control strain positive HM-1:IMSS. Last 9 belonged to symptomatic patients with pathogenic strain.
These results indicate that ethnic amplified by polymerase chain reaction is insufficiently to establish differential diagnostic. Therefore is necessary carry out enzyme digestion to identify pathogenic strain.
溶组织内阿米巴的半胱氨酸蛋白酶被认为是阿米巴病发病机制中的重要毒力因子。基于编码30 kDa蛋白酶的种族基因差异。本研究验证了一种通过限制性图谱区分致病性和非致病性溶组织内阿米巴菌株的策略。
使用来自4名无症状和9名有症状患者的13份含有溶组织内阿米巴囊肿的粪便样本,患者年龄和性别不同,接种于罗宾逊培养基。获得的DNA用于扩增种族基因,并用限制性内切酶Taq I和Hinf I进行切割。
所有菌株均在罗宾逊培养基中培养。获得了一个与溶组织内阿米巴探针杂交的530 bp片段。通过用Taq I和Hinf I进行限制性图谱评估表明,4名无症状患者中的2份样本属于致病菌株。这与对照菌株阳性HM-1:IMSS一致。其余9份属于有症状的致病菌株患者。
这些结果表明,通过聚合酶链反应扩增的种族基因不足以进行鉴别诊断。因此,有必要进行酶切以鉴定致病菌株。