scholarly journals Detection A2058G and A2059G on the 23S rRNA Gene by Multiplex Nested PCR to Identify Treponema pallidum Resistance to Azithromycin in Indonesia

2021 ◽  
Author(s):  
Yeva Rosana ◽  
Andi Yasmon ◽  
Wresti Indriatmi ◽  
Ida Effendi ◽  
Raden Lia Kusumawati ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Treponema Pallidum ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene

scholarly journals Macrolide treatment failure in a case of secondary syphilis: a novel A2059G mutation in the 23S rRNA gene of Treponema pallidum subsp. pallidum

2009 ◽  
Vol 58 (6) ◽  
pp. 832-836 ◽  
Author(s):  
Petra Matějková ◽  
Magdalena Flasarová ◽  
Hana Zákoucká ◽  
Milan Bořek ◽  
Soňa Křemenová ◽  
...  
Keyword(s):  
Treatment Failure ◽  
Molecular Detection ◽  
Treponema Pallidum ◽  
Secondary Syphilis ◽  
23S Rrna ◽  
Rrna Gene ◽  
The Czech Republic ◽  
Clinical Specimens ◽  
Gene Position ◽  
23S Rrna Gene

We report an occurrence of treatment failure after oral spiramycin therapy in a man with secondary syphilis and a reported penicillin and tetracycline allergy. Molecular detection revealed treponemal DNA in the blood of the patient and sequencing of the 23S rDNA identified an A to G transition at the gene position corresponding to position 2059 in the Escherichia coli 23S rRNA gene. The occurrence of this novel 23S rDNA mutation was examined among 7 rabbit-propagated syphilitic strains of Treponema pallidum and among 22 syphilis patient isolates from the Czech Republic. The prevalence of A2058G and A2059G mutations among clinical specimens was 18.2 and 18.2 %, respectively.

scholarly journals Detection and identification of azithromycin resistance mutations on Treponema pallidum 23S rRNA gene by nested multiplex polymerase chain reaction

2017 ◽  
Vol 26 (2) ◽  
pp. 90-6 ◽  
Author(s):  
Desy A. Gultom ◽  
Yeva Rosana ◽  
Ida Efendi ◽  
Wresti Indriatmi ◽  
Andi Yasmon
Keyword(s):  
Multiplex Polymerase Chain Reaction ◽  
Treponema Pallidum ◽  
23S Rrna ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Detection And Identification ◽  
23S Rrna Gene ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Blood Specimens

Background: 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.

scholarly journals Molecular Typing and Macrolide Resistance Analyses ofTreponema pallidumin Heterosexuals and Men Who Have Sex with Men in Japan, 2017

2018 ◽  
Vol 57 (1) ◽  
Author(s):  
Mizue Kanai ◽  
Yuzo Arima ◽  
Shingo Nishiki ◽  
Ken Shimuta ◽  
Ichiro Itoda ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Treponema Pallidum ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
Content Type ◽  
Heterosexual Women ◽  
23S Rrna Gene ◽  
Strain Type ◽  
Sex With Men

ABSTRACTIn recent years, syphilis notifications have increased dramatically in Japan. We carried out molecular typing and macrolide resistance analyses ofTreponema pallidumsubsp.pallidumsamples collected from patients at four clinics and a hospital in Tokyo and Osaka prefectures in 2017. The macrolide resistant strain type 14d/f (SS14-like clade) was found in significantly more cases of syphilis among heterosexuals than in those among men who have sex with men (MSM); i.e., 79% (31/39) of the strains from heterosexuals were 14d/f compared to 37% (7/19) of those from MSM (odds ratio [OR], 6.6; 95% confidence interval [CI], 1.7 to 26.7;P = 0.002). In addition, 83% (50/60) of the strains were identified as macrolide resistant with an A2058G mutation in the 23S rRNA gene; 90% (35/39) of the strains from heterosexuals were macrolide resistant compared to 58% (11/19) of those from MSM. The odds of having the resistant mutation were considerably higher in the former (OR, 6.4; 95% CI, 1.3 to 33.5;P = 0.02). Heterosexual women and heterosexual men showed similar distributions, and the association remained the same when restricted to men. The strain type distribution and the prevalence of macrolide resistance differed substantially between syphilis strains from heterosexual cases and from MSM cases, suggesting distinct epidemiologic profiles for the two communities and providing important insight into the dynamics of syphilis in Japan.

scholarly journals Detection of mixed clarithromycin-resistant and -susceptible Helicobacter pylori using nested PCR and direct sequencing of DNA extracted from faeces

2007 ◽  
Vol 56 (9) ◽  
pp. 1174-1180 ◽  
Author(s):  
Norihisa Noguchi ◽  
Emiko Rimbara ◽  
Ayami Kato ◽  
Akifumi Tanaka ◽  
Kengo Tokunaga ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Nested Pcr ◽  
Direct Sequencing ◽  
Accurate Method ◽  
23S Rrna ◽  
Rrna Gene ◽  
Peptic Ulcers ◽  
Unsuccessful Treatment ◽  
23S Rrna Gene ◽  
H Pylori

The major cause of chemotherapy failure in patients with chronic gastritis and peptic ulcers caused by Helicobacter pylori is clarithromycin (CAM) resistance due to a mutation in the 23S rRNA gene. This study describes a non-invasive and accurate method for the detection of mixed CAM-resistant and -susceptible H. pylori by sequencing of the H. pylori 23S rRNA gene. Faeces were crushed with beads and the 23S rRNA gene was amplified using a nested PCR on the extracted DNA. Mutation analysis of this gene using this method showed that 20.4 % of patients carried mixed CAM-susceptible (wild type) and -resistant (A2142G or A2143G mutant) H. pylori. Furthermore, it was found that 66.6 % of patients who had been treated unsuccessfully carried one of these mutations in the 23S rRNA gene (including the mixed type), whilst standard culture detected CAM-resistant isolates in only 22.2 % of patients with unsuccessful treatment. These data suggest that, for successful therapy, the diagnosis method described here would more accurately detect CAM-resistant H. pylori, including mixed infections.

scholarly journals Molecular typing and macrolide resistance analyses ofTreponema pallidumin heterosexuals and men who have sex with men communities in Japan, 2017

2018 ◽  
Author(s):  
Mizue Kanai ◽  
Yuzo Arima ◽  
Shingo Nishiki ◽  
Ken Shimuta ◽  
Ichiro Itoda ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Treponema Pallidum ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene ◽  
Strain Type ◽  
Sex With Men ◽  
Insight Into ◽  
Resistant Mutation

AbstractIn recent years, syphilis notifications have increased dramatically in Japan. We performed molecular typing and analyzed macrolide resistance ofTreponema pallidumsamples collected from four clinics and a hospital in Tokyo and Osaka prefectures in 2017.Macrolide resistant strain type 14d/f was found significantly more in heterosexual syphilis cases compared with those strains identified in men who have sex with men (MSM) syphilis cases. The proportion of 14d/f among heterosexuals was 79% (31/39) compared to 37% (7/19) among MSM [OR, 6.6 (95% CI, 1.7 to 26.7); P=0.002]. 83% (50/60) of the strains were identified as macrolide resistant with an A2058G mutation at the 23S rRNA gene. 90% (35/39) of the heterosexual strains were macrolide resistant, relative to 58% (11/19) of MSM strains; the odds of having the resistant mutation was considerably higher in heterosexuals compared with MSM [OR, 6.4 (95% CI, 1.3 to 33.5); P=0.02]. Heterosexual females and males showed similar distributions, and the results remained the same when restricted to men.The strain type distribution and frequency of macrolide resistance differed substantially between heterosexual and MSM syphilis samples, suggesting distinct epidemiologic profiles for the two communities and insight into syphilis transmission dynamics in Japan.

scholarly journals Panel of 23S rRNA Gene-Based Real-Time PCR Assays for Improved Universal and Group-Specific Detection of Phytoplasmas

2009 ◽  
Vol 75 (9) ◽  
pp. 2945-2950 ◽  
Author(s):  
Jennifer Hodgetts ◽  
Neil Boonham ◽  
Rick Mumford ◽  
Matthew Dickinson
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Multiplex Assay ◽  
The Other ◽  
23S Rrna ◽  
Rrna Gene ◽  
Specific Detection ◽  
23S Rrna Gene ◽  
Pcr Assays

ABSTRACT Primers and probes based on the 23S rRNA gene have been utilized to design a range of real-time PCR assays for routine phytoplasma diagnostics. These assays have been authenticated as phytoplasma specific and shown to be at least as sensitive as nested PCR. A universal assay to detect all phytoplasmas has been developed, along with a multiplex assay to discriminate 16SrI group phytoplasmas from members of all of the other 16Sr groups. Assays for the 16SrII, 16SrIV, and 16SrXII groups have also been developed to confirm that the 23S rRNA gene can be used to design group-specific assays.

First Report of the 23S rRNA Gene A2058G Point Mutation Associated With Macrolide Resistance in Treponema pallidum From Syphilis Patients in Cuba

2016 ◽  
Vol 43 (5) ◽  
pp. 332-334 ◽  
Author(s):  
Angel A. Noda ◽  
Nelvis Matos ◽  
Orestes Blanco ◽  
Islay Rodríguez ◽  
Lola Virginia Stamm
Keyword(s):  
Point Mutation ◽  
Treponema Pallidum ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
First Report ◽  
23S Rrna Gene

scholarly journals Molecular pathways to high-level azithromycin resistance in Neisseria gonorrhoeae

2021 ◽  
Author(s):  
J G E Laumen ◽  
S S Manoharan-Basil ◽  
E Verhoeven ◽  
S Abdellati ◽  
I De Baetselier ◽  
...  
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Ribosomal Proteins ◽  
Efflux Pump ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene ◽  
Evolution Of Resistance ◽  
High Level ◽  
Azithromycin Resistance ◽  
Level Resistance

Abstract Background The prevalence of azithromycin resistance in Neisseria gonorrhoeae is increasing in numerous populations worldwide. Objectives To characterize the genetic pathways leading to high-level azithromycin resistance. Methods A customized morbidostat was used to subject two N. gonorrhoeae reference strains (WHO-F and WHO-X) to dynamically sustained azithromycin pressure. We tracked stepwise evolution of resistance by whole genome sequencing. Results Within 26 days, all cultures evolved high-level azithromycin resistance. Typically, the first step towards resistance was found in transitory mutations in genes rplD, rplV and rpmH (encoding the ribosomal proteins L4, L22 and L34 respectively), followed by mutations in the MtrCDE-encoded efflux pump and the 23S rRNA gene. Low- to high-level resistance was associated with mutations in the ribosomal proteins and MtrCDE efflux pump. However, high-level resistance was consistently associated with mutations in the 23S ribosomal RNA, mainly the well-known A2059G and C2611T mutations, but also at position A2058G. Conclusions This study enabled us to track previously reported mutations and identify novel mutations in ribosomal proteins (L4, L22 and L34) that may play a role in the genesis of azithromycin resistance in N. gonorrhoeae.

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