Detection of A2058G and A2059G on the 23S rRNA Gene by Multiplex Nested PCR to Identify <i>Treponema pallidum</i> Resistance to Azithromycin in Indonesia

Abstract

Azithromycin is one of the antibiotics used to treat syphilis, especially in the context of penicillin allergy. Resistance to azithromycin is widely reported associated with one and/or two point mutations on the 23S rRNA gene but has yet to be described in Indonesia.Specimens were collected from 220 patients diagnosed with secondary syphilis. A multiplex nested PCR testing system using the 23S rRNA target gene of Treponema pallidum was designed using three pairs of primers. The first step used PCR pairs of primers to detect T. pallidum. In the second step of PCR using 2 pairs of primers were achieved to identify azithromycin resistant T. pallidum based on A2058G and A2059G point mutations. There was no T.pallidum identified resistant to azithromycin in Jakarta and Bandung. T. pallidum resistance to azithromycin were found in Makassar, Medan, and Bali. The majority of azithromycin resistance was found among heterosexual males and in patients living with HIV. This study has demonstrated T. pallidum resistance to azithromycin in Indonesia appears to be a novel variant of resistance, containing both the A2058G and A2059G mutations that was found in Medan and Makassar.

Molecular Characteristics of Macrolide Resistance in Treponema pallidum from Patients with Latent Syphilis in Xinjiang, China

“…Thus, the detection rate of T. pallidum could be improved by using an optimal PCR approach and increasing the number of specific target genes. 7,20 Nevertheless, among 27 T. pallidum-positive samples, the A2058G mutation was identified in 24 samples (88.9%) (Figure 1D) and the A2059G mutation was found in three samples (11.1%) (Figure 1E). These findings indicate that macrolide resistance in T. pallidum exists in patients with latent syphilis in Xinjiang, and the A2058G mutation is the predominant mechanism of macrolide resistance in T. pallidum, which is consistent with the results reported in previous studies in Shanghai (95.4%), 8 Hunan (97.5%), 9 Shangdong (92.1%), 10 and Xiamen (100%).…”

“…Thus, the detection rate of T. pallidum could be improved by using an optimal PCR approach and increasing the number of specific target genes. 7 , 20 …”

“…4,5 Previous studies indicated that A2058G mutation and A2059G mutation in the 23S rRNA gene of T. pallidum are common mechanisms causing macrolide resistance in T. pallidum. 6,7 In China, high rates of A2058G mutation have been reported in Shanghai (95.4%), 8 Hunan (97.5%), 9 Shangdong (92.1%), 10 and Xiamen (100%), 11 whereas the A2059G mutation in the 23S rRNA gene of T. pallidum has only been reported in Shangdong, China, by Li et al 10 Taking swab samples from chancres was the most common method used to isolate T. pallidum for the detection of A2058G and A2059G point mutations in the 23S rRNA gene of T. pallidum in previous studies. [12][13][14] However, it is impossible to isolate T. pallidum from chancres in patients with latent syphilis, who have an absence of chancres and the other syphilis skin lesions.…”