Spiroplasma

Moyaba, Percy, Ouedraogo, Gisele Ms, Pagabeleguem, Soumaïla, Njokou, Flobert, Freitas, Ngambia, Vreysen, Marc Jb & Abd-Alla, Adly Mm, 2023, Prevalence of Spiroplasma and interaction with wild Glossina tachinoides microbiota, Parasite (Paris, France) 30 (62), pp. 1-14 : 4

Genetic variation and phylogenetic analysis of Spiroplasma in G. tachinoides

To assess the genetic variation of Spiroplasma in wild G. tachinoides , an MLST approach was employed on positive samples from Burkina Faso and Ghana using the following genes: 16S rRNA, Spiroplasma fructose repressor (fruR), Spiroplasma DNA Topoisomerase 4 subunit B (parE), and RNA polymerase subunit beta (rpoB). Primer sets used for each reaction, product sizes, and PCR conditions are shown in Supplementary Table 2.

All amplified PCR products were purified using a High Pure PCR Cleanup Micro Kit (Roche Diagnostics, Indianapolis, IN, USA) and a ZR-96 DNA Clean-up Kit™ (Zymo Research, Irvine, CA, USA). Sequencing was performed with Eurofins Genomics Company (https://www.eurofinsgenomics.com) and sequencing data were first analyzed using Geneious Prime ® 2023.0.2 and then blasted using the “Blast” resource of NCBI to confirm them as Spiroplasma . Phylogenetic trees were built for each gene (16S rRNA, fruR, pare, and rpoB) and for the concatenated data set using all four gene sequences. Multiple alignments were then performed using MUSCLE alignment with the default parameters on Geneious Prime ® 2023.0.2 and the Neighbor-joining tree was built using the Tamura-Nei genetic distance model.