Polymerase chain reaction confirmation of diagnosis of intestinal amebiasis in Puducherry.

Background Stool microscopy, the commonly used test for diagnosis of intestinal amebiasis, is unreliable as it does not differentiate Entamoeba histolytica, the causative agent for amebiasis, from non-pathogenic Entamoeba dispar and Entamoeba moshkovskii. Methods Two hundred and forty-six stool samples collected between January to February 2009, were examined microscopically for E. histolytica/E. dispar/E. moshkovskii complex cysts and trophozoites. Nested multiplex PCR targeting a 16 S-like rRNA gene for differential detection of all the three morphologically similar forms of E. histolytica, E. moshkovskii and E. dispar simultaneously was done on all microscopy positive stool samples. Results Forty-nine stool samples were positive for Entamoeba spp. by microscopy. Nested multiplex PCR for E. histolytica, E. moshkovskii and E. dispar was positive in only 19 of these samples, and only 6 of the 19 samples were positive for E. histolytica. Conclusion Stool microscopy grossly over-diagnosed intestinal amebiasis and is likely to result in unnecessary treatment with anti-protozoal drugs.

Detection of Entamoeba histolytica DNA in the saliva of amoebic liver abscess patients who received prior treatment with metronidazole.

Saliva is an easily-accessible and a non-invasive clinical specimen alternate to blood and liver pus. An attempt was made to detect Entamoeba histolytica DNA released in the saliva of amoebic liver abscess (ALA) patients by applying 16S-like rRNA gene-based nested multiplex polymerase chain reaction (NM-PCR). The NM-PCR detected E. histolytica DNA in the saliva of eight (28.6%) of 28 ALA patients. The NM-PCR result was negative for E. histolytica DNA in the saliva of all the eight ALA patients who were tested prior to treatment with metronidazole but was positive in the saliva of eight (40%) of 20 ALA patient who were tested after therapy with metronidazole. The NM-PCR detected E. histolytica DNA in liver abscess pus of all 28 (100%) patients with ALA. The TechLab E. histolytica II enzyme-linked immunosorbent assay was positive for E. histolytica Gal/GalNAc lectin antigen in the liver abscess pus of 13 (46.4%) of the 28 ALA patients. The indirect haemagglutination (IHA) test was positive for anti-amoebic antibodies in the serum of 22 (78.6%) of the 28 ALA patients and 2 (5.7%) of 35 healthy controls. The present study, for the first time, demonstrates the release of E. histolytica DNA in the saliva of ALA patients by applying NM-PCR.

Entamoeba moshkovskii and Entamoeba dispar-associated infections in pondicherry, India.

The prevalence of Laredo strain--Entamoeba moshkovskii--and non-pathogenic E. dispar in patients attending the Jawaharlal Institute of Postgraduate Medical Education and Research hospital, Pondicherry, India, is reported here. E. moshkovskii is reported for the first time in India. The species are morphologically indistinguishable from pathogenic E. histolytica. Of 746 stool samples screened, 68 showing cyst or trophozoite stage of E. histolytica, E. dispar, or E. moshkovskii were subjected to small subunit (SSU) rRNA gene-based polymerase chain reaction, which revealed a higher prevalence of E. dispar (8.8%) and E. moshkovskii (2.2%) compared to E. histolytica (1.7%) in patients. Only 19% of the 68 stool samples, resembling E. histolytica by microscopy, were actually E. histolytica, implying that 81% of suspected infections were misdiagnosed and would have been treated unnecessarily with anti-amoebic drugs.