Detection of excretory Entamoeba histolytica DNA in the urine, and detection of E. histolytica DNA and lectin antigen in the liver abscess pus for the diagnosis of amoebic liver abscess.
BACKGROUND
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.


RESULTS
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.


CONCLUSIONS
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.
Parija
Subhash C
SC
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India. parijasc@vsnl.com
Khairnar
Krishna
K
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000032:analysis*; D000981:Antiprotozoal Agents / Q000008:administration & dosage / Q000627:therapeutic use; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis / Q000302:isolation & purification / Q000652:urine*; D004748:Entamoeba histolytica / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004749:Entamoebiasis / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology; D004797:Enzyme-Linked Immunosorbent Assay; D006386:Hemagglutination Tests; D006801:Humans; D037102:Lectins / Q000032:analysis; D008101:Liver Abscess, Amebic / Q000175:diagnosis* / Q000188:drug therapy / Q000469:parasitology; D008795:Metronidazole / Q000008:administration & dosage / Q000627:therapeutic use; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013492:Suppuration / Q000276:immunology / Q000469:parasitology*
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000981:Antiprotozoal Agents; D017931:DNA Primers; D016054:DNA, Protozoan; D037102:Lectins; D012336:RNA, Ribosomal, 16S; D008795:Metronidazole