Giardia duodenalis

K, Pamela C., oster, Lapuente, Juan, Pizarro, Andrea, Prieto-P, Laura, erez, Ramon, erez-Tanoira, Dashti, Alejandro, Begona ˜ Bailo, Muadica, Aly S., Gonz, David, alez-Barrio, Calero-Bernal, Rafael, Ponce-Gordo, Francisco & Carmena, David, 2022, Presence and genetic diversity of enteric protists in captive and semi-captive non-human primates in coteˆd’Ivoire, Sierra Leone, and Peru, International Journal for Parasitology: Parasites and Wildlife 17, pp. 26-34 : 28

2.8. Molecular detection and characterization of Giardia duodenalis

We conducted G. duodenalis DNA detection using a real-time PCR (qPCR) method targeting a 62-bp region of the gene codifying the ssu rRNA gene of the parasite ( Verweij et al., 2003b). Amplification reactions (25 μL) consisted of 3 μL of template DNA, 0.5 μM of each primer Gd-80F and Gd-127R, 0.4 μM of probe ( Table S1), and 12.5 μL TaqMan® Gene Expression Master Mix (Applied Biosystems). Cycling conditions and data analysis were as described above for the detection of E. histolytica / E. dispar .

We subsequently assessed G. duodenalis isolates that tested positive by qPCR by sequence-based multi-locus genotyping of the genes encoding for the glutamate dehydrogenase (gdh) ( Read et al., 2004), β- giardin (bg) ( Lalle et al., 2005), and triose phosphate (tpi) ( Sulaiman et al., 2003) proteins of the parasite. We conducted amplifications by semi-nested and nested PCR protocols using specific primer pairs ( Table S1).