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Journal of Clinical Microbiology, May 2005, p. 2168-2172, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2168-2172.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Real-Time-PCR Assay for Diagnosis of Entamoeba histolytica Infection

Shantanu Roy,1 Mamun Kabir,1 Dinesh Mondal,1 Ibne Karim M. Ali,1,2 William A. Petri Jr.,3* and Rashidul Haque1

International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), Dhaka, Bangladesh,1 London School of Hygiene and Tropical Medicine, London, England,2 Departments of Medicine, Microbiology, and Pathology, University of Virginia, Charlottesville, Virginia3

Received 18 October 2004/ Returned for modification 8 November 2004/ Accepted 20 December 2004

We developed a real-time-PCR assay utilizing a molecular-beacon probe for the detection of Entamoeba histolytica and compared its sensitivity to stool antigen detection and traditional PCR. A total of 205 stool and liver abscess pus specimens from patients and controls were used for this purpose, 101 (49%) of which were positive by the TechLab E. histolytica-specific antigen detection test, while the other 104 (51%) stool and liver abscess pus specimens were negative by the antigen detection test. DNA was extracted from the stool and liver abscess pus specimens by the QIAGEN method and the small-subunit rRNA gene of E. histolytica and then amplified by traditional and real-time PCR. Out of these 205 stool and liver abscess pus specimens, 124 were positive by the real-time-PCR assay and 90 were positive by the traditional-PCR test. Compared to the real-time-PCR assay, the antigen detection test was 79% sensitive and 96% specific. When the traditional-PCR test results were compared to the real-time-PCR assay, the sensitivity of traditional PCR was 72% and the specificity was 99%. In conclusion, all three methods for the detection of E. histolytica were highly specific, with real-time PCR being the most sensitive.

* Corresponding author. Mailing address: Division of Infectious Diseases and International Health, P.O. Box 801340, Rm. 2115, MR4 Building, University of Virginia Health System, Charlottesville, VA 22908-1340. Phone: (434) 924-5621. Fax: (434) 924-0075. E-mail: wap3g{at}virginia.edu.

Journal of Clinical Microbiology, May 2005, p. 2168-2172, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2168-2172.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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