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Table 4 Primers and probes for detection and sequencing in this study

From: Decontamination of mycoplasma-contaminated Orientia tsutsugamushistrains by repeating passages through cell cultures with antibiotics

Targets Assay Name Primers and probes
Mycoplasmas    
tuf genea) real-time PCR Mollicutes 414F 5'-TCCAGGWCAYGCTGACTA-3'
   Mollicutes 541R 5'-ATTTTWGGAACKCCWACTTG-3'
   Probe 451Fa) 5'-GGTGCTGCACAAATGGATGG-3'
tuf gene Sequencing 1st Myco-tuf-F1 5'-HATHGGCCAYRTTGAYCAYGGKAAAA-3'
   Myco-tuf-F2 5'-ATGATYACHGGDGCWGCHCAAATGGA-3'
  Sequencing 2nd Myco-tuf-R1 5'-CCRCCTTCRCGRATDGAGAAYTT-3'
   Myco-tuf-R2 5'-TKTRTGACGDCCACCTTCYTC-3'
16s-23s rRNA intergenic spacer region nested PCR 1st MCGpF11 5'-ACACCATGGGAGYTGGTAAT-3'
   R23-1R 5'-CTCCTAGTGCCAAGSCATYC-3'
  nested PCR 2nd R16-2 5'-GTGSGGMTGGATCACCTCCT-3'
   MCGpR21 5'-GCATCCACCAWAWACYCTT-3'
Orientia tsutsugamushi    
47kDa common antigen coding gene real-time PCR Ots-47k-F 5'-AATTCGTCGTGGTATGTTAAATG-3'
   Ots-47k-R 5'-AGCAATTCCACATTGTGCTG-3'
   Ots-47k-P b) 5'-TGCTTAATGAATTAACTCCAGAATT-3'
  1. a) Locked nucleic acid (LNA) bases (underlined) and was synthesized with the fluorescent reporter 6-carboxyfluorescein (FAM) covalently coupled to the 5’ end and a dark quencher to the 3’ end.
  2. b) TaqMan probe was synthesized with the fluorescent reporter 6-carboxyfluorescein (FAM) covalently coupled to the 5’ end and a dark quencher to the 3’ end.