scholarly journals Macrolide Susceptibility and Molecular Characteristics of Bordetella Pertussis

2021 ◽  
Author(s):  
juansheng zhang ◽  
diqiang zhang ◽  
xiaoqiang wang ◽  
xiaoguang wei ◽  
hao li ◽  
...  
Keyword(s):  
Bordetella Pertussis ◽  
Variable Number Tandem Repeat ◽  
Resistance Mutation ◽  
Drug Resistance Mutation ◽  
Genetic Mutation ◽  
Variable Number ◽  
Macrolide Resistance ◽  
Rrna Genes ◽  
23S Rrna ◽  
Molecular Characteristics

Abstract ObjectiveTo compare the macrolide resistance and molecular characteristics of clinical isolated Bordetella pertussis, and explore the relationship between the macrolide-resistance and genotypes. MethodsErythromycin、azithromycin and clarithromycin susceptibility of clinical isolates during 2018-2020 was determined by E-test. The A2047G of the 23S rRNA genes was sequenced for drug-resistance mutation. Multilocus antigen sequence typing (MAST)、Multiple Locus Variable-number tandem repeat Analysis (MLVA) and Pulsed field gel electrophoresis (PFGE) methods were employed to do molecular typing for the strains. Results58 strains were isolated in this study, 46 of them were macrolide-resistant and 12 sensitive. All macrolide-resistant strains carried a genetic mutation at the A2047G site, genotype was prn1/ptxP1/ptxA1/fim3-1/fim2-1, the MLVA types were identified as MT195、MT55 and MT104, PFGE profiles were classified into BPSR23 and BpFINR9 types. However, no mutations were found in all macrolide-sensitive strains whose genotypes were (prn9 or prn2)/ptxP1/ptxA1/fim3-1/fim2-1 and MT27, and PFGE classified other profiles. ConclusionsThe clinical isolated Bordetella pertussis has serious resistance to erythromycin and began to spread to other macrolides. There were differences between macrolide-resistant and -sensitive Bordetella pertussis in genotypes. The acquisition of macrolide resistance may be associated with change of specific molecular types.

scholarly journals Macrolide resistance and molecular typing of Mycoplasma pneumoniae infections during a 4 year period in Spain

2020 ◽  
Vol 75 (10) ◽  
pp. 2752-2759 ◽  
Author(s):  
Belén Rivaya ◽  
Elena Jordana-Lluch ◽  
Gema Fernández-Rivas ◽  
Sònia Molinos ◽  
Roi Campos ◽  
...  
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Variable Number Tandem Repeat ◽  
Respiratory Viruses ◽  
Variable Number ◽  
Community Acquired Pneumonia ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Resistance Pattern ◽  
Molecular Tools ◽  
Resistance Detection

Abstract Background Mycoplasma pneumoniae (MP) causes community-acquired pneumonia affecting mainly children, and tends to produce cyclic outbreaks. The widespread use of macrolides is increasing resistance rates to these antibiotics. Molecular tools can help in diagnosis, typing and resistance detection, leading to better patient management. Objectives To assess the MP genotypes and resistance pattern circulating in our area while comparing serological and molecular diagnosis of MP. Methods Molecular and serological diagnosis of MP was performed in 821 samples collected in Badalona (Barcelona, Spain) from 2013 to 2017. Multiple locus variable number tandem repeat analysis (MLVA) and macrolide resistance detection by pyrosequencing were performed in those cases positive by PCR. Presence of respiratory viruses and relevant clinical data were also recorded. Results MP was detected in 16.8% of cases by PCR, with an overall agreement with serology of 76%. Eleven different MLVA types were identified, with 4-5-7-2 (50.1%) and 3-5-6-2 (29.2%) being the most abundant, with the latter showing a seasonal increase during the study. A total of 8% of the strains harboured a point substitution associated with macrolide resistance, corresponding mainly to an A2063G 23S rRNA mutation and directly related to previous macrolide therapy. Analysis of respiratory viruses showed viral coinfections in most cases. Conclusions Serological and molecular tools combined could improve MP diagnosis and the analysis of its infection patterns. Macrolide resistance is associated with previous therapy. Given that MP pneumonia usually resolves spontaneously, it should be reconsidered whether antibiotic treatment is suitable for all cases.

scholarly journals Multiple-Locus Variable-Number Tandem Repeat Analysis of Dutch Bordetella pertussis Strains Reveals Rapid Genetic Changes with Clonal Expansion during the Late 1990s

2004 ◽  
Vol 186 (16) ◽  
pp. 5496-5505 ◽  
Author(s):  
Leo M. Schouls ◽  
Han G. J. van der Heide ◽  
Luc Vauterin ◽  
Paul Vauterin ◽  
Frits R. Mooi
Keyword(s):  
The Netherlands ◽  
Tandem Repeat ◽  
Bordetella Pertussis ◽  
Minimum Spanning Tree ◽  
Whooping Cough ◽  
Variable Number Tandem Repeat ◽  
Clonal Expansion ◽  
Variable Number ◽  
Multiple Locus ◽  
Repeat Analysis

ABSTRACT Bordetella pertussis, the causative agent of whooping cough, has remained endemic in The Netherlands despite extensive nationwide vaccination since 1953. In the 1990s, several epidemic periods have resulted in many cases of pertussis. We have proposed that strain variation has played a major role in the upsurges of this disease in The Netherlands. Therefore, molecular characterization of strains is important in identifying the causes of pertussis epidemiology. For this reason, we have developed a multiple-locus variable-number tandem repeat analysis (MLVA) typing system for B. pertussis. By combining the MLVA profile with the allelic profile based on multiple-antigen sequence typing, we were able to further differentiate strains. The relationships between the various genotypes were visualized by constructing a minimum spanning tree. MLVA of Dutch strains of B. pertussis revealed that the genotypes of the strains isolated in the prevaccination period were diverse and clearly distinct from the strains isolated in the 1990s. Furthermore, there was a decrease in diversity in the strains from the late 1990s, with a remarkable clonal expansion that coincided with the epidemic periods. Using this genotyping, we have been able to show that B. pertussis is much more dynamic than expected.

scholarly journals Macrolide resistance in Helicobacter pylori: rapid detection of point mutations and assays of macrolide binding to ribosomes.

1997 ◽  
Vol 41 (12) ◽  
pp. 2724-2728 ◽  
Author(s):  
A Occhialini ◽  
M Urdaci ◽  
F Doucet-Populaire ◽  
C M Bébéar ◽  
H Lamouliatte ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Point Mutations ◽  
Restriction Enzymes ◽  
The United States ◽  
Drug Binding ◽  
Macrolide Resistance ◽  
Rrna Genes ◽  
23S Rrna ◽  
Dependent Manner ◽  
H Pylori

Resistance of Helicobacter pylori to macrolides is a major cause of failure of eradication therapies. Single base substitutions in the H. pylori 23S rRNA genes have been associated with macrolide resistance in the United States. Our goal was to extend this work to European strains, to determine the consequence of this mutation on erythromycin binding to H. pylori ribosomes, and to find a quick method to detect the mutation. Seven pairs of H. pylori strains were used, the parent strain being naturally susceptible to macrolides and the second strain having acquired an in vivo resistance during a treatment regimen that included clarithromycin. The identity of the strains was confirmed by random amplified polymorphic DNA testing with two different primers, indicating that resistance was the result of the selection of variants of the infecting strain. All resistant strains were found to have point mutations at position 2143 (three cases) or 2144 (four cases) but never on the opposite DNA fragment of domain V of the 23S rRNA gene. The mutation was A-->G in all cases except one (A-->C) at position 2143. Using BsaI and BbsI restriction enzymes on the amplified products, we confirmed the mutations of A-->G at positions 2144 and 2143, respectively. Macrolide binding was tested on purified ribosomes isolated from four pairs of strains with [14C]erythromycin. Erythromycin binding increased in a dose-dependent manner for the susceptible strain but not for the resistant one. In conclusion we suggest that the limited disruption of the peptidyltransferase loop conformation, caused by a point mutation, reduces drug binding and consequently confers resistance to macrolides. Finally, the macrolide resistance could be detected without sequencing by performing restriction fragment length polymorphism with appropriate restriction enzymes.

scholarly journals 23S rRNA base pair 2057-2611 determines ketolide susceptibility and fitness cost of the macrolide resistance mutation 2058A->G

2005 ◽  
Vol 102 (14) ◽  
pp. 5180-5185 ◽  
Author(s):  
P. Pfister ◽  
N. Corti ◽  
S. Hobbie ◽  
C. Bruell ◽  
R. Zarivach ◽  
...  
Keyword(s):  
Base Pair ◽  
Resistance Mutation ◽  
Macrolide Resistance ◽  
Fitness Cost ◽  
23S Rrna

scholarly journals Spontaneous Erythromycin Resistance Mutation in a 23S rRNA Gene, rrlA, of the Extreme Thermophile Thermus thermophilus IB-21

2001 ◽  
Vol 183 (14) ◽  
pp. 4382-4385 ◽  
Author(s):  
Steven T. Gregory ◽  
Jamie H. D. Cate ◽  
Albert E. Dahlberg
Keyword(s):  
Genetic Manipulation ◽  
Thermus Thermophilus ◽  
Resistance Mutation ◽  
Rrna Genes ◽  
23S Rrna ◽  
Rrna Gene ◽  
Erythromycin Resistance ◽  
23S Rrna Gene ◽  
Resistant Mutants ◽  
23S Rrna Genes

ABSTRACT Spontaneous, erythromycin-resistant mutants of Thermus thermophilus IB-21 were isolated and found to carry the mutation A2058G in one of two 23S rRNA operons. The heterozygosity of these mutants indicates that A2058G confers a dominant or codominant phenotype in this organism. This mutation provides a valuable tool for the genetic manipulation of the 23S rRNA genes ofThermus.

scholarly journals Heterogeneous Macrolide Resistance and Gene Conversion in the Pneumococcus

2006 ◽  
Vol 50 (1) ◽  
pp. 359-361 ◽  
Author(s):  
Nicole Wolter ◽  
Anthony M. Smith ◽  
David J. Farrell ◽  
Keith P. Klugman
Keyword(s):  
Streptococcus Pneumoniae ◽  
Gene Conversion ◽  
Clinical Isolate ◽  
Resistant Strain ◽  
Macrolide Resistance ◽  
Fitness Cost ◽  
Rrna Genes ◽  
23S Rrna ◽  
Resistant Phenotype ◽  
23S Rrna Genes

ABSTRACT A macrolide-resistant clinical isolate of Streptococcus pneumoniae with 23S rRNA mutations showed a heterogeneous phenotype and genotype. The mutant 23S rRNA genes from this isolate transformed susceptible strain R6 to resistance. Culture of resistant strain R6 in the absence of antibiotic pressure showed gene conversion to occur between the four 23S rRNA alleles, resulting in reversion to susceptibility with the resistant phenotype showing a fitness cost. These data explain the disappearance on subculture of heterogeneous macrolide resistance in the pneumococcus.

scholarly journals Mycoplasma genitalium in Symptomatic Male Urethritis: Macrolide Use Is Associated With Increased Resistance

2019 ◽  
Vol 70 (5) ◽  
pp. 805-810 ◽  
Author(s):  
Yang Li ◽  
Xiaohong Su ◽  
Wenjing Le ◽  
Sai Li ◽  
Zhaoyan Yang ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Mycoplasma Genitalium ◽  
Macrolide Resistance ◽  
Rrna Genes ◽  
Fluoroquinolone Resistance ◽  
23S Rrna ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
23S Ribosomal Rna ◽  
The Impact

Abstract Background Mycoplasma genitalium (MG) causes symptomatic urethritis in men, and can infect alone or together with other sexually transmitted infection (STI) agents. Methods The prevalence of MG and other STIs was determined in 1816 men with symptomatic urethritis. Resistance of MG to macrolides and fluoroquinolones was determined by sequencing; the impact of recent antimicrobial usage on the distribution of MG single or mixed infections was determined. Results Overall, prevalence of MG infection was 19.7% (358/1816). Fifty-four percent (166/307) of MG infections occurred alone in the absence of other STI agents. Men with single MG infection self-administered or were prescribed antibiotics more often in the 30 days prior to enrollment than subjects with urethritis caused by MG coinfection (P < .0001). Higher rates (96.7%) of infection with macrolide resistance in MG were identified in men who had taken macrolides prior to enrollment (P < .03). Overall, 88.9% (303/341) of 23S ribosomal RNA (rRNA) genes contained mutations responsible for macrolide resistance; 89.5% (308/344) of parC and 12.4% (42/339) of gyrA genes had mutations responsible for fluoroquinolone resistance. Approximately 88% (270/308) of MG had combined mutations in 23S rRNA and parC genes; 10.4% (32/308) had mutations in all 3 genes. Conclusions MG was the single pathogen identified in 11% of men with symptomatic urethritis. Overall, nearly 90% of MG infections were resistant to macrolides and fluoroquinolones. Men who took macrolides in the 30 days prior to enrollment had higher rates (97%) of macrolide-resistant MG. Resistance was associated with numerous mutations in 23SrRNA, parC, and gyrA genes.

scholarly journals First Description of Macrolide-Resistant Mycoplasma pneumoniae in Adults with Community-Acquired Pneumonia in Italy

2019 ◽  
Vol 2019 ◽  
pp. 1-5 ◽  
Author(s):  
Daniela Loconsole ◽  
Anna Lisa De Robertis ◽  
Rosanna Mallamaci ◽  
Anna Sallustio ◽  
Anna Morea ◽  
...  
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Variable Number Tandem Repeat ◽  
Point Mutations ◽  
Cross Sectional Study ◽  
Variable Number ◽  
Community Acquired Pneumonia ◽  
23S Rrna ◽  
Cross Sectional ◽  
South Italy ◽  
Multiple Loci

Background. Mycoplasma pneumoniae is a common cause of community-acquired pneumonia (CAP). This cross-sectional study aimed to determine the prevalence of macrolide-resistant M. pneumoniae strains in a convenience series of 234 adult hospitalised and nonhospitalised subjects with a diagnosis of CAP in January 2013 to April 2015 in South Italy. Methods. Respiratory samples were subjected to real-time PCR. In M. pneumoniae-positive samples, domain V of 23S rRNA was sequenced to detect resistance-conferring point mutations. P1 major adhesion protein typing and multiple loci variable-number tandem repeat analysis (MLVA) were also performed. Results. Of the 234 samples, 15 (6.4%) were positive for M. pneumoniae. Three of these had a macrolide-resistant genotype: two and one had A2063G and A2064G mutations, respectively. Fourteen of the 15 strains were subtyped: half had subtype 1 and half had subtype 2. Eight strains underwent MLVA profiling: one each had the J, A, and Z type. The remainder was unclassifiable. Conclusions. This novel discovery of macrolide-resistant M. pneumoniae strains in adults with CAP in Italy suggests that there may be increasing circulation of these strains in the population. To facilitate rapid optimization of the antibiotic strategy in Italy, macrolide resistance should be monitored by a surveillance system that is based on molecular methods.

scholarly journals Rapid and simple SNP genotyping for Bordetella pertussis epidemic strain MT27 based on a multiplexed single-base extension assay

2020 ◽  
Author(s):  
Kazunari Kamachi ◽  
Shu-Man Yao ◽  
Chuen-Sheue Chiang ◽  
Kentaro Koide ◽  
Nao Otsuka ◽  
...  
Keyword(s):  
Bordetella Pertussis ◽  
Diversity Index ◽  
Temporal Trends ◽  
Variable Number Tandem Repeat ◽  
Genotypic Diversity ◽  
Variable Number ◽  
Snp Genotyping ◽  
Single Base Extension ◽  
Single Base ◽  
Base Extension

Abstract Multilocus variable-number tandem repeat analysis (MLVA) is widely used for genotyping of Bordetella pertussis, the causative bacteria for pertussis. However, MLVA genotyping is losing its discriminate power because prevalence of the epidemic MT27 strain (MLVA-27) is increasing worldwide. To address this, we developed a single nucleotide polymorphism (SNP) genotyping method for MT27 based on multiplexed single-base extension (SBE) assay. A total of 237 MT27 isolates collected in Japan during 1999−2018 were genotyped and classified into ten SNP genotypes (SG1 to SG10) with a Simpson’s diversity index (DI) of 0.79 (95% CI, 0.76–0.82). Temporal trends showed a marked increase in the genotypic diversity in the 2010s: Simpson’s DI was zero in 1999–2004, 0.16 in 2005–2009, 0.83 in 2010–2014, and 0.76 in 2015–2018. This indicates that the SNP genotyping is applicable to the recently circulating MT27 strain. Additionally, almost all outbreak-associated MT27 isolates were classified into the same SNP genotypes for each outbreak. Multiplexed SBE assay allows for rapid and simple genotyping, indicating that the SNP genotyping can potentially be a useful tool for subtyping the B. pertussis MT27 strain in routine surveillance and outbreak investigations.

Sign in / Sign up

Export Citation Format

Share Document