Detection of entamoeba histolytica dna in the saliva of amoebic liver abscess patients who received prior treatment with metronidazole

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J HEALTH POPUL NUTR 2008 Dec;26(4):418-425 ISSN 1606-0997 | $ 5.00+0.20

©INTERNATIONAL CENTRE FOR DIARRHOEAL DISEASE RESEARCH, BANGLADESH

Detection of Entamoeba histolytica DNA in the Saliva of Amoebic Liver Abscess Patients Who Received Prior Treatment with Metronidazole
Krishna Khairnar and Subhash Chandra Parija
Department of Microbiology, Jawaharlal Institute of Postgraduate MedicalEducation and Research, Puducherry 605 006, India

ABSTRACT
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. Key words: Amoebiasis; DNA; Entamoeba histolytica; Metronidazole; Polymerase chain reaction; Saliva; India

INTRODUCTION
The use of saliva as a diagnostic fluid has been increasingly reported worldwide in the last decade. Technological advancement has taken place during the pastfew years enabling the use of saliva as a clinical specimen to diagnose disease and predict disease progression (1). Initially, saliva was used as a clinical specimen for antibody detection in the diagnosis of infectious diseases. Detection of salivary antibody was found to be useful for the diagnosis of bacterial infections caused by Helicobacter pylori, Shigella, and Borrelia burgdorferi (2-4)and various viral infections, such as hepatitis A, hepatitis B, hepatitis C, measles,
Correspondence and reprint requests should be addressed to: Dr. Subhash Chandra Parija Director-Professor and Head Department of Microbiology Jawaharlal Institute of Postgraduate Medical Education and Research Puducherry 605 006 India Email: parijasc@vsnl.com Fax: 91-413-2272067

mumps, rubella, rotavirus,dengue, parvovirus B 19, and HIV (5-11). Detection of salivary antibody has also been studied for the diagnosis of some parasitic infections caused by Toxoplasma gondii, Schistosoma mansoni, Taenia solium, and Entamoeba histolytica (12-15). Subsequently, saliva has also been used for the detection of antigen in the diagnosis of pneumococcal pneumonia (16), hepatitis B virus, measles, mumps, and rubella(17-20). There is only one report till date on the detection of salivary lectin antigen of E. histolytica for the diagnosis of amoebic liver abscess (ALA) with a sensitivity and specificity of 22% and 97.4% respectively (21). The reports on the use of saliva for the detection of DNA for the diagnosis of infectious diseases, however, are limited (22-26). The polymerase chain reaction (PCR) hasbeen used for facilitating diagnosis of viral infections, such as Epstein-Barr, cytomegalovirus, human herpes virus 6, 7, and 8, and rabies using saliva (22-25). The PCR has also been evaluated for the detection of H. pylori-associated peptic ulcer, by demonstration of H. pylori DNA in saliva (26). However, reports on the detection of DNA

Detection of salivary Entamoeba histolytica DNA...
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