Parija Subhash C, Khairnar Krishna
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.
BMC Microbiol. 2007 May 18;7:41. doi: 10.1186/1471-2180-7-41.
Amoebic liver abscess (ALA) and pyogenic liver abscesses (PLA) appear identical by ultrasound and other imaging techniques. Collection of blood or liver abscess pus for diagnosis of liver abscesses is an invasive procedure, and the procedure requires technical expertise and disposable syringes. Collection of urine is a noninvasive procedure. Therefore, there has been much interest shown towards the use of urine as an alternative clinical specimen for the diagnosis of some parasitic infections. Here, we report for the first time the detection of E. histolytica DNA excreted in the urine for diagnosis of the cases of ALA.
E. histolytica DNA was detected in liver abscess pus specimen of 80.4% of ALA patients by a nested multiplex polymerase chain reaction (PCR) targeting 16S-like r RNA gene. The nested PCR detected E. histolytica DNA in all 37 (100%) liver abscess pus specimens collected prior to metronidazole treatment, but were detected in only 53 of 75 (70.6%) pus specimens collected after therapy with metronidazole. Similarly, the PCR detected E. histolytica DNA in 21 of 53 (39.6%) urine specimens of ALA patients. The test detected E. histolytica DNA in only 4 of 23 (17.4%) urine specimens collected prior to metronidazole treatment, but were detected in 17 of 30 (56.7%) urine specimens collected after treatment with metronidazole. The enzyme-linked immunosorbent assay (ELISA) for the detection of lectin E. histolytica antigen in the liver abscess pus showed a sensitivity of 50% and the indirect haemagglutination (IHA) test for detection of amoebic antibodies in the serum showed a sensitivity of 76.8% for the diagnosis of the ALA.
The present study for the first time shows that the kidney barrier in ALA patients is permeable to E. histolytica DNA molecule resulting in excretion of E. histolytica DNA in urine which can be detected by PCR. The study also shows that the PCR for detection of E. histolytica DNA in urine of patients with ALA can also be used as a prognostic marker to assess the course of the diseases following therapy by metronidazole. The detection of E. histolytica DNA in urine specimen of ALA patients provides a new approach for the diagnosis of ALA.
通过超声和其他成像技术,阿米巴肝脓肿(ALA)和化脓性肝脓肿(PLA)看起来相同。采集血液或肝脓肿脓液用于诊断肝脓肿是一种侵入性操作,该操作需要专业技术和一次性注射器。采集尿液则是一种非侵入性操作。因此,人们对将尿液用作诊断某些寄生虫感染的替代临床标本表现出了浓厚兴趣。在此,我们首次报告了通过检测尿液中排泄的溶组织内阿米巴DNA来诊断ALA病例。
通过针对16S样rRNA基因的巢式多重聚合酶链反应(PCR),在80.4%的ALA患者的肝脓肿脓液标本中检测到了溶组织内阿米巴DNA。巢式PCR在所有37份(100%)在甲硝唑治疗前采集的肝脓肿脓液标本中检测到了溶组织内阿米巴DNA,但在甲硝唑治疗后采集的75份脓液标本中仅在53份(70.6%)中检测到。同样,PCR在53份ALA患者的尿液标本中的21份(39.6%)中检测到了溶组织内阿米巴DNA。该检测在甲硝唑治疗前采集的23份尿液标本中仅在4份(17.4%)中检测到溶组织内阿米巴DNA,但在甲硝唑治疗后采集的30份尿液标本中的I7份(56.7%)中检测到。用于检测肝脓肿脓液中溶组织内阿米巴凝集素抗原的酶联免疫吸附测定(ELISA)显示敏感性为50%,用于检测血清中阿米巴抗体的间接血凝试验(IHA)对ALA诊断的敏感性为76.8%。
本研究首次表明,ALA患者的肾脏屏障对溶组织内阿米巴DNA分子具有通透性,导致尿液中排泄溶组织内阿米巴DNA,可通过PCR检测到。该研究还表明,检测ALA患者尿液中溶组织内阿米巴DNA的PCR也可作为一种预后标志物,用于评估甲硝唑治疗后疾病的进程。检测ALA患者尿液标本中的溶组织内阿米巴DNA为ALA的诊断提供了一种新方法。