Impact of population structure in the design of RNA-based diagnostics for antibiotic resistance in <i>Neisseria gonorrhoeae</i>

Cb, Wadsworth; Mr, Sater; Rp, Bhattacharyya; Yh, Grad

doi:10.1101/537175

Cited by 2 publications

(3 citation statements)

References 43 publications

(60 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The transcriptional profiles of the CRO RS isolates were similar to one another but 126 distinct from GCGS0457 (Supplementary Figure 2), with 1337 and 1410 annotated ORFs 127 significantly altered in abundance, respectively. The transcriptomic differences we observe may 128 partly reflect the genetic distance between these isolates 16 , as the resistant strains GCGS1095 129 and GCGS1014 are more closely related to one another than they are to GCGS0457. We 130 therefore compared GCGS0457 to its isogenic CRO RS transformants GCGS0457 RpoB R201H and 131 GCGS0457 RpoD E98K .…”

Section: Results and Discussion 34mentioning

confidence: 86%

RNA polymerase mutations cause cephalosporin resistance in clinicalNeisseria gonorrhoeaeisolates

Palace

Wang

Rubin

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

1The rising incidence of Neisseria gonorrhoeae infection and antimicrobial resistance 2 imperils effective therapy for gonococcal infections 1-4 . Current first-line therapy for gonorrhea 3 relies on ceftriaxone (CRO), an extended spectrum cephalosporin (ESC), as the backbone of 4 treatment. Dual therapy with azithromycin was intended to delay the emergence of resistance, 5 but rising azithromycin resistance rates have been reported 2,5 , resulting in revision of United 6Kingdom treatment guidelines to CRO monotherapy 6 . With no clear next-line agent, gonococcal 7 resistance to ESCs is a problem of paramount clinical importance, underscored by recent reports 8 of treatment failure and global spread of a multidrug-resistant strain 4,7 . Reduced susceptibility to 9 ceftriaxone (CRO RS ) is most commonly associated with interspecies mosaic and other alleles of 10 penA (PBP2) 8-10 , the primary target of ESCs 8,11 . However, collections of clinical specimens 11 often include isolates with high-level reduced susceptibility to ESCs (ESC RS ) that is not 12 attributable to penA variation 3,12,13 . Here, we describe the genetic basis of reduced susceptibility 13 in three such isolates collected in the United States. Each of these isolates has a unique mutation 14 in the RNA polymerase holoenzyme that causes phenotypic CRO RS when introduced into 15 susceptible strains. We demonstrate that other clinical isolates can develop high-level ESC RS via 16 a single nucleotide change in RNA polymerase. This result has important implications regarding 17 the biology underlying cephalosporin resistance, the potential for de novo evolution of resistance 18 under cephalosporin monotherapy, and the accuracy of sequence-based diagnostics. 19

show abstract

Section: Results and Discussion 34mentioning

confidence: 86%

RNA polymerase mutations cause cephalosporin resistance in clinicalNeisseria gonorrhoeaeisolates

Palace

Wang

Rubin

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Our analysis shows that expression levels of antibiotic target genes are significantly higher in susceptible isolates compared to nonsusceptible isolates. A recent RNA-based study identified candidate markers from the transcriptomes which were highly expressed in the susceptible isolates only [51].…”

Section: Discussionmentioning

confidence: 99%

“…A range of molecular diagnostic approaches have been evaluated, each with its strengths and limitations [42,[47][48][49][50][51]. While most methods target specific mutations to infer resistance, our approach targets the gene and its expression levels to infer resistance.…”

Section: Introductionmentioning

confidence: 99%

Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis

Mitchev

Singh

Ramsuran

et al. 2022

International Journal of Microbiology

View full text Add to dashboard Cite

Introduction. Treatment of gonorrhoea infection is limited by the increasing prevalence of multidrug-resistant strains. Cost-effective molecular diagnostic tests can guide effective antimicrobial stewardship. The aim of this study was to correlate mRNA expression levels in Neisseria gonorrhoeae antibiotic target genes and efflux pump genes to antibiotic resistance in our population. Methods. This study investigated the expression profile of antibiotic resistance-associated genes (penA, ponA, pilQ, mtrR, mtrA, mtrF, gyrA, parC, parE, rpsJ, 16S rRNA, and 23S rRNA) and efflux pump genes (macAB, norM, and mtrCDE), by quantitative real-time PCR, in clinical isolates from KwaZulu-Natal, South Africa. Whole-genome sequencing was used to determine the presence or absence of mutations. Results. N. gonorrhoeae isolates, from female and male patients presenting for care at clinics in KwaZulu-Natal, South Africa, were analysed. As determined by binomial regression and ROC analysis, the most significant ( p ≤ 0.05 ) markers for resistance prediction in this population, and their cutoff values, were determined to be mtrC ( p = 0.024 ; cutoff <0.089), gyrA ( p = 0.027 ; cutoff <0.0518), parE ( p = 0.036 ; cutoff <0.0033), rpsJ ( p = 0.047 ; cutoff <0.0012), and 23S rRNA ( p = 0.042 ; cutoff >7.754). Conclusion. Antimicrobial stewardship includes exploring options to conserve currently available drugs for gonorrhoea treatment. There is the potential to predict an isolate as either susceptible or nonsusceptible based on the mRNA expression level of specific candidate markers, to inform patient management. This real-time qPCR approach, with few targets, can be further investigated for use as a potentially cost-effective diagnostic tool to detect resistance.

show abstract

Impact of population structure in the design of RNA-based diagnostics for antibiotic resistance in Neisseria gonorrhoeae

Cited by 2 publications

References 43 publications

RNA polymerase mutations cause cephalosporin resistance in clinicalNeisseria gonorrhoeaeisolates

RNA polymerase mutations cause cephalosporin resistance in clinicalNeisseria gonorrhoeaeisolates

Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis

Contact Info

Product

Resources

About