Journal
KOREAN JOURNAL OF PARASITOLOGY
Volume 54, Issue 5, Pages 631-636Publisher
KOREAN SOC PARASITOLOGY, SEOUL NATL UNIV COLL MEDI
DOI: 10.3347/kjp.2016.54.5.631
Keywords
Cryptosporidium parvum; Giardia lamblia; Cyclospora cayetanensis; multiplex PCR; touchdown PCR; stool sample
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Funding
- Korea Centers for Disease Control and Prevention, Republic of Korea [2015-E54003-00]
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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 > 1 x 10(3) oocysts for C. parvum, > 1 x 10(4) cysts for G. lamblia, and > 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.
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