Multiplex polymerase chain reaction method to detect Cyclospora, Cystoisospora, and Microsporidia in stool samples

作者：Taniuchi Mami^*; Verweij Jaco J; Sethabutr Orntipa; Bodhidatta Lada****; Garcia Lynne; Maro Athanasia; Kumburu Happiness; Gratz Jean; Kibiki Gibson; Houpt Eric R

来源：Diagnostic Microbiology and Infectious Disease, 2011, 71(4): 386-390.

DOI：10.1016/j.diagmicrobio.2011.08.012

• PCR
• Microsporidia
• Cystoisospora belli
• Cystoisospora
• Isospora belli
• Isospora
• Diarrhea
• Enteropathogens
• Encephalitozoon intestinalis
• Enterocytozoon bieneusi
• Cyclospora cayetanensis
• Cyclospora
• Multiplex PCR
• Fecal PCR
• Stool PCR
• Luminex

Cyclospora, Cystoisospora, and Microsporidia are eukaryotic enteropathogens that are difficult to detect in stool samples because they require special stains and microscopy. We developed a multiplex polymerase chain reaction (PCR) reaction with 4 primer sets to amplify Cyclospora cayetanensis, Cystoisospora belli, Enterocytozoon bieneusi, and Encephalitozoon intestinalis. Detection of the amplicon is through specific probes coupled to Luminex beads. Sensitivity of the assay was evaluated using Encephalitozoon intestinalis spores and revealed detection of 10(1) spores spiked into stool. No cross-reactivity was observed. We evaluated the assay on diarrheal specimens from Thailand, Tanzania, Indonesia, and the Netherlands that had been previously tested by microscopy, and the assay yielded 87-100% sensitivity and 88-100% specificity. Microscopy-negative/PCR-positive samples had lower Luminex values, suggesting they were true but with lower burden infections. In summary, this is a convenient single PCR reaction that can detect Cyclospora, Cystoisospora, and Microsporidia without the need for cumbersome microscopic analysis.

• 出版日期2011-12
• 单位河北医科大学