Korean J Parasitol.  2011 Sep;49(3):281-284. 10.3347/kjp.2011.49.3.281.

PCR Diagnosis of Entamoeba histolytica Cysts in Stool Samples

• Moon JH
• Cho SH
• Yu JR
• Lee WJ
• Cheun HI

• KMID: 1715652
• DOI: http://doi.org/10.3347/kjp.2011.49.3.281

Abstract

Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.

Keyword

Entamoeba histolytica; SSU rDNA; PCR; diagnosis; stool

MeSH Terms

DNA Primers/genetics
DNA, Protozoan/genetics
DNA, Ribosomal/genetics
Entamoeba histolytica/genetics/*isolation & purification
Entamoebiasis/*diagnosis
Histones/genetics
Humans
Molecular Diagnostic Techniques/*methods
Parasitology/*methods
Polymerase Chain Reaction/*methods
Protozoan Proteins/genetics
Sensitivity and Specificity