Evaluation of a new single-tube multiprobe real-time PCR for diagnosis of entamoeba histolytica and entamoeba dispar

Shih Yu Liang, Kan Tai Hsia, Yun Hsien Chan, Chia Kwung Fan, Donald Dah Shyong Jiang, Olfert Landt, Dar Der Ji

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10 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)793-797
Number of pages5
JournalJournal of Parasitology
Issue number4
Publication statusPublished - Aug 2010


ASJC Scopus subject areas

  • Parasitology
  • Ecology, Evolution, Behavior and Systematics

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