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1 August 2010 Evaluation of a New Single-Tube Multiprobe Real-Time PCR for Diagnosis of Entamoeba histolytica and Entamoeba dispar
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Abstract
A single-tube multiprobe real-time PCR assay for simultaneous detection of Entamoeba histolytica and Entamoeba dispar was developed. One primer pair with 2 species-specific probes was designed based on new SSU RNA regions of the ribosomal DNA-containing episome. The sensitivity is 1 parasite per milliliter of feces and thus superior to the conventional nested PCR and comparable to other published real-time PCR protocols. The applicability for clinical diagnosis was validated with 218 stool specimens from patients. A total of 51 E. histolytica and 39 E. dispar positive samples was detected by the multiprobe real-time PCR compared to 39 and 22 by routine nested PCR diagnosis. The detection rate of Entamoeba species for the multiprobe real-time PCR assays was significantly higher than the nested PCR (40.8% vs. 28.0%, P < 0.01). The test did not show cross reactivity with DNA from Entamoeba moshkovskii, Giardia lamblia, Cryptosporidium sp., Escherichia coli, or other nonpathogenic enteric parasites. The multiprobe real-time PCR assay is simple and rapid and has high specificity and sensitivity. The assay could streamline the laboratory diagnosis procedure and facilitate epidemiological investigation.
Shih-Yu Liang, Kan-Tai Hsia, Yun-Hsien Chan, Chia-Kwung Fan, Donald Dah-Shyong Jiang, Olfert Landt and Dar-Der Ji "Evaluation of a New Single-Tube Multiprobe Real-Time PCR for Diagnosis of Entamoeba histolytica and Entamoeba dispar," Journal of Parasitology 96(4), (1 August 2010). https://doi.org/10.1645/GE-2373.1
Received: 1 November 2009; Accepted: 1 March 2010; Published: 1 August 2010
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