Detection and identification of azithromycin resistance mutations on Treponema pallidum 23S rRNA gene by nested multiplex polymerase chain reaction
Abstract:
Azithromycin-resistant strains of Treponema pallidum is associated with the
mutation of 23S rRNA gene of T. pallidum. Although these strains are now
prevalent in many countries, there is no laboratory test kit to detect and
identify these mutations. Thus, in this study we developed a nested multiplex
polymerase chain reaction (PCR) to detect and identify A2058G and A2059G
mutations in 23S rRNA gene.
Methods: Three primer sets were designed for nested PCR reactions. To
obtain maximum PCR reaction, all parameters were optimized. The specificity of
the primer sets was evaluated towards some microorganisms. A sensitivity test
was conducted to get detection limit of deoxyribonucleic acid (DNA). Forty-five
whole blood specimens were tested by PCR, and positive results were confirmed
by the DNA sequencing.
Results: The assay could detect at least 4,400 DNA copy number and showed
no cross reaction with other microorganisms used in the specificity test. A
total 13 of 45 whole blood specimens were PCR positive for T. pallidum, and no
single mutations (either A2058G or A2059G) were detected. Two positive
specimens were confirmed by the DNA sequencing and showed no mutation.
Conclusion: Nested multiplex PCR developed in this study showed a
specific and sensitive test for the detection and identification of A2058G
and/or A2059G mutations of 23S rRNA T. pallidum gene.
Keywords: PCR; syphilis;
Treponema pallidum resistance
Author: Desy A. Gultom, Yeva
Rosana, Ida Efendi, Wresti Indriatmi, Andi Yasmon
Journal Code: jpkedokterangg170202
