School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, Malaysia.
BMC Infect Dis. 2013 Mar 21;13:144. doi: 10.1186/1471-2334-13-144.
Amoebic liver abscess (ALA) is the most frequent clinical presentation of extra-intestinal amoebiasis. The diagnosis of ALA is typically based on the developing clinical symptoms, characteristic changes on radiological imaging and serology. Numerous serological tests have been introduced for the diagnosis of ALA, either detecting circulating amoebic antigens or antibodies. However those tests show some pitfalls in their efficacy and/or the preparation of the tests are costly and tedious. The commercial IHA kit that used crude antigen was reported to be useful in diagnosis of ALA, however high antibody background in endemic areas may cause problems in its interpretation. Thus, discovery of well-defined antigen(s) is urgently needed to improve the weaknesses of current serodiagnostic tests.
Crude antigen of E. histolytica was analysed by 2-DE and Western blot to identify a protein of diagnostic potential for ALA. The corresponding gene of the antigenic protein was then cloned, expressed and the purified recombinant protein was subsequently evaluated for serodiagnosis of ALA in an indirect ELISA format.
Analysis of crude antigen showed that phosphoglucomutase (PGM) has the diagnostic potential. Recombinant PGM (rPGM) showed 79.17% (19/24) sensitivity and 86.67% (195/225) specificity in diagnosis of ALA based on the COV of mean +1SD. There was no significant difference between rPGM-ELISA and IHA diagnostic kit in the diagnosis of ALA in terms of sensitivity and specificity at p-value < 0.05.
In conclusion, rPGM-ELISA is found to be useful for serodiagnosis of ALA. Future studies will determine whether rPGM-ELISA also detects antibodies produced in amoebic dysentery and asymptomatic cases.
阿米巴肝脓肿(ALA)是肠外阿米巴病最常见的临床表现。ALA 的诊断通常基于临床症状的发展、影像学和血清学的特征性变化。已经引入了许多血清学检测方法来诊断 ALA,这些方法可以检测循环中的阿米巴抗原或抗体。然而,这些检测方法在其有效性方面存在一些缺陷,或者检测方法的准备既昂贵又繁琐。据报道,使用粗抗原的商业 IHA 试剂盒可用于诊断 ALA,但在流行地区,高抗体背景可能会导致其解释出现问题。因此,迫切需要发现明确的抗原,以改善当前血清学诊断检测的弱点。
使用 2-DE 和 Western blot 分析溶组织内阿米巴的粗抗原,以鉴定对 ALA 具有诊断潜力的蛋白质。然后克隆相应的抗原蛋白基因,表达并纯化重组蛋白,随后在间接 ELISA 格式中评估其对 ALA 的血清学诊断。
分析粗抗原表明磷酸葡萄糖变位酶(PGM)具有诊断潜力。重组 PGM(rPGM)在基于均值+1SD 的 COV 下,对 ALA 的诊断显示出 79.17%(19/24)的敏感性和 86.67%(195/225)的特异性。在敏感性和特异性方面,rPGM-ELISA 与 IHA 诊断试剂盒在 ALA 的诊断中没有显著差异,p 值<0.05。
总之,rPGM-ELISA 被发现对 ALA 的血清学诊断有用。未来的研究将确定 rPGM-ELISA 是否也能检测出在阿米巴痢疾和无症状病例中产生的抗体。