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 >1×103 oocysts for C. parvum, >1×104 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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A novel TaqMan probe-based pentaplex qPCR assay for the simultaneous detection of five pathogenic protozoans Zheng-qin Gao, Rui Fu Scientific Reports.2025;[Epub] CrossRef
A Multiplex RT-PCR for the Detection of Three Viruses and One Viroid Infecting Hemp Derrick J. Grunwald, Jacob MacWilliams, Laine Hackenberg, Sydney Stroschein, Renee Rioux, Punya Nachappa, Shelby Ellison PhytoFrontiers™.2024; 4(4): 524. CrossRef
Efficacy of a membrane concentration method combined with real-time PCR for detection of Giardia and Cryptosporidium in drinking water Jiang Jingyi, Yao Ping, Xu Jian, Chen Jia, Mao Xujian, Li Qiong, Tu Bowen, Wang Fengming Letters in Applied Microbiology.2023;[Epub] CrossRef
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Sporadic cyclosporiasis in symptomatic Cuban patients: Confirmation of positive results from conventional diagnostic methods by molecular assay Luis Enrique Jerez Puebla, Fidel A. Núñez Fernández, Jorge Fraga Nodarse, Iraís Atencio Millán, Iredys Cruz Rodríguez, Lisette Pérez Santos, Yanet Pintos Saavedra, Laura Rodríguez Moreno, Lucy J. Robertson Diagnostic Microbiology and Infectious Disease.2020; 97(3): 115048. CrossRef
Human cyclosporiasis Annunziata Giangaspero, Robin B Gasser The Lancet Infectious Diseases.2019; 19(7): e226. CrossRef
Cyclospora cayetanensis and Cyclosporiasis: An Update Sonia Almeria, Hediye N. Cinar, Jitender P. Dubey Microorganisms.2019; 7(9): 317. CrossRef
Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples Estelle Menu, Charles Mary, Isabelle Toga, Didier Raoult, Stéphane Ranque, Fadi Bittar BMC Research Notes.2018;[Epub] CrossRef
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Entamoeba histolytica: an Overview Nadia A. El-Dib Current Tropical Medicine Reports.2017; 4(1): 11. CrossRef
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