Multiplex-Touchdown PCR to Simultaneously Detect , , and , the Major Causes of Traveler's Diarrhea.

This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., , , and , the major causes of traveler's diarrhea and are resistant to standard antimicrobial treatments. The target genes included the oocyst wall protein for , Glutamate dehydrogenase for , and 18S ribosomal RNA (18S rRNA) for . The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10 oocysts for , >1×10 cysts for , and >1 copy of the 18S rRNA gene for . Taken together, our protocol convincingly demonstrated the ability to simultaneously detect , , and in stool samples.