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This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum , Giardia lamblia , and Cyclospora cayetanensis , the major causes of traveler's diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum , Glutamate dehydrogenase for G. lamblia , and 18S ribosomal RNA (18S rRNA) for C. cayetanensis . 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 &gt;1×10 3 oocysts for C. parvum , &gt;1×10 4 cysts for G. lamblia , and &gt;1 copy of the 18S rRNA gene for C. cayetanensis . Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum , G. lamblia , and C. cayetanenesis in stool samples.

korean journal of parasitology

Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler’s Diarrhea