Molecular Assay for Detection of Genetic Markers Associated with Decreased Susceptibility to Cephalosporins in Neisseria gonorrhoeae.

Abstract:

:The incidence of antimicrobial-resistant Neisseria gonorrhoeae continues to rise in Canada; however, antimicrobial resistance data are lacking for approximately 70% of gonorrhea infections that are diagnosed directly from clinical specimens by nucleic acid amplification tests (NAATs). We developed a molecular assay for surveillance use to detect mutations in genes associated with decreased susceptibility to cephalosporins that can be applied to both culture isolates and clinical samples. Real-time PCR assays were developed to detect single nucleotide polymorphisms (SNPs) in ponA, mtrR, penA, porB, and one N. gonorrhoeae-specific marker (porA). We tested the real-time PCR assay with 252 gonococcal isolates, 50 nongonococcal isolates, 24 N. gonorrhoeae-negative NAAT specimens, and 34 N. gonorrhoeae-positive NAAT specimens. Twenty-four of the N. gonorrhoeae-positive NAAT specimens had matched culture isolates. Assay results were confirmed by comparison with whole-genome sequencing data. For 252 N. gonorrhoeae strains, the agreement between the DNA sequence and real-time PCR was 100% for porA, ponA, and penA, 99.6% for mtrR, and 95.2% for porB. The presence of ≥2 SNPs correlated with decreased susceptibility to ceftriaxone (sensitivities of >98%) and cefixime (sensitivities of >96%). Of 24 NAAT specimens with matched cultures, the agreement between the DNA sequence and real-time PCR was 100% for porB, 95.8% for ponA and mtrR, and 91.7% for penA. We demonstrated the utility of a real-time PCR assay for sensitive detection of known markers for the decreased susceptibility to cephalosporins in N. gonorrhoeae. Preliminary results with clinical NAAT specimens were also promising, as they correlated well with bacterial culture results.

journal_name

J Clin Microbiol

authors

Peterson SW,Martin I,Demczuk W,Bharat A,Hoang L,Wylie J,Allen V,Lefebvre B,Tyrrell G,Horsman G,Haldane D,Garceau R,Wong T,Mulvey MR

doi

10.1128/JCM.00493-15

subject

Has Abstract

pub_date

2015-07-01 00:00:00

pages

2042-8

issue

7

eissn

0095-1137

issn

1098-660X

pii

JCM.00493-15

journal_volume

53

pub_type

杂志文章
  • Routine identification of Mycobacterium tuberculosis complex isolates by automated hybridization.

    abstract::Methodologies for biochemical identification of mycobacteria isolated from clinical samples are still cumbersome, taking skilled technicians 3 to 6 weeks. We describe here a 2-h identification system for mycobacterial isolates belonging to the Mycobacterium tuberculosis complex using a DNA probe. After 30 min of hands...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.32.11.2702-2705.1994

    authors: Mabilat C,Desvarenne S,Panteix G,Machabert N,Bernillon MH,Guardiola G,Cros P

    更新日期:1994-11-01 00:00:00

  • Performance of the Vitek MS v2.0 system in distinguishing Streptococcus pneumoniae from nonpneumococcal species of the Streptococcus mitis group.

    abstract::The Vitek MS v2.0 matrix-assisted laser desorption ionization-time of flight mass spectrometry system accurately distinguished Streptococcus pneumoniae from nonpneumococcal S. mitis group species. Only 1 of 116 nonpneumococcal isolates (<1%) was misidentified as S. pneumoniae. None of 95 pneumococcal isolates was misi...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00824-13

    authors: Branda JA,Markham RP,Garner CD,Rychert JA,Ferraro MJ

    更新日期:2013-09-01 00:00:00

  • Identification of nocardiae and streptomycetes of medical importance.

    abstract::During a taxonomic investigation of Nocardia spp. and Streptomyces spp., we received 658 isolates from laboratories of both human and veterinary medicine. Our procedure leading to the identification of 92% of these isolates, the species that they represented, and a characteristic pattern of properties of the strains o...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.11.6.728-736.1980

    authors: Mishra SK,Gordon RE,Barnett DA

    更新日期:1980-06-01 00:00:00

  • Geographical heterogeneity between Far Eastern and Western countries in prevalence of the virulence plasmid, the superantigen Yersinia pseudotuberculosis-derived mitogen, and the high-pathogenicity island among Yersinia pseudotuberculosis strains.

    abstract::Yersinia pseudotuberculosis produces novel superantigenic toxins designated YPMa (Y. pseudotuberculosis-derived mitogen), YPMb, and YPMc and has a pathogenicity island termed HPI (high-pathogenicity island) and R-HPI (the right-hand part of the HPI with truncation in its left-hand part) on the chromosome. Analysis of ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.39.10.3541-3547.2001

    authors: Fukushima H,Matsuda Y,Seki R,Tsubokura M,Takeda N,Shubin FN,Paik IK,Zheng XB

    更新日期:2001-10-01 00:00:00

  • Detection of cytomegalovirus antibody with latex agglutination.

    abstract::Transfusion-acquired cytomegalovirus (CMV) infections should be prevented in seronegative immunocompromised patients by providing blood products from donors who are also seronegative. Latex agglutination was investigated as a simple and rapid method for detecting antibody against CMV. Latex beads were coated with CMV ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.22.1.68-70.1985

    authors: Adler SP,McVoy M,Biro VG,Britt WJ,Hider P,Marshall D

    更新日期:1985-07-01 00:00:00

  • Detection and sequence analysis of Danish and Swedish strains of mink astrovirus.

    abstract::The sequences of mink astroviruses collected from 11 farms in Denmark and Sweden were analyzed and found to be homologous with one another but different from those of other astroviruses. A species-specific reverse transcriptase-PCR for mink astrovirus was established and shown to be suitable for the analysis of clinic...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.11.5192-5194.2003

    authors: Mittelholzer C,Englund L,Hedlund KO,Dietz HH,Svensson L

    更新日期:2003-11-01 00:00:00

  • Update on the major clonal types of methicillin-resistant Staphylococcus aureus in the Czech Republic.

    abstract::The purpose of our study was the molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in 21 hospitals in the Czech Republic in the period 2000-2002 and comparison with previous results from 1996-1997. Strains were analyzed by pulsed-field gel electrophoresis (PFGE) of SmaI ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.11.4998-5005.2003

    authors: Melter O,Aires de Sousa M,Urbásková P,Jakubů V,Zemlicková H,de Lencastre H

    更新日期:2003-11-01 00:00:00

  • Rapid identification and fingerprinting of Candida krusei by PCR-based amplification of the species-specific repetitive polymorphic sequence CKRS-1.

    abstract::A PCR method was developed to identify and fingerprint Candida krusei isolates simply and rapidly. The primer pair Arno1 and Arno2 was designed to amplify the polymorphic species-specific repetitive sequence CKRS-1 (C. krusei repeated sequence 1) that we identified in the nontranscribed intergenic regions (IGRs) of rR...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.35.6.1337-1343.1997

    authors: Carlotti A,Chaib F,Couble A,Bourgeois N,Blanchard V,Villard J

    更新日期:1997-06-01 00:00:00

  • Detection and Prevalence of Penicillin-Susceptible Staphylococcus aureus in the United States in 2013.

    abstract::Using blaZ PCR as the "gold standard," the sensitivities of CLSI penicillin zone edge and nitrocefin-based tests for β-lactamase production in Staphylococcus aureus were 64.5% and 35.5%, respectively, with specificity of 99.8% for both methods. In 2013, 13.5% of 3,083 S. aureus isolates from 31 U.S. centers were penic...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.03109-15

    authors: Richter SS,Doern GV,Heilmann KP,Miner S,Tendolkar S,Riahi F,Diekema DJ

    更新日期:2016-03-01 00:00:00

  • Comparison of BD Bactec Plus blood culture media to VersaTREK Redox blood culture media for detection of bacterial pathogens in simulated adult blood cultures containing therapeutic concentrations of antibiotics.

    abstract::Antibiotic neutralization in blood culture media from two automated systems was evaluated by measuring the recovery of organisms and times to detection in simulated cultures. Overall, BD Bactec Plus media (Bactec FX system) outperformed TREK 80 ml Redox media (VersaTREK system), although results suggest a relative rat...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01958-10

    authors: Miller NS,Rogan D,Orr BL,Whitney D

    更新日期:2011-04-01 00:00:00

  • PCR for detection and identification of Abiotrophia spp.

    abstract::Members of the genus Abiotrophia, formerly known as nutritionally variant streptococci, are important pathogens causing septicemia and endocarditis. Cultivation and biochemical differentiation of Abiotrophia spp. are often difficult. Based on 16S rRNA sequences, two PCR assays for detection and identification of Abiot...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.36.10.2844-2846.1998

    authors: Roggenkamp A,Leitritz L,Baus K,Falsen E,Heesemann J

    更新日期:1998-10-01 00:00:00

  • Discrimination of antibody to herpes B virus from antibody to herpes simplex virus types 1 and 2 in human and macaque sera.

    abstract::The antigenic cross-reactive characteristics of herpes B virus and herpes simplex virus (HSV) type 1 (HSV-1) and HSV-2 are responsible for false-positive diagnoses by serological assays in humans and macaques. In the present study, we developed a fluorometric indirect enzyme-linked immunosorbent assay (ELISA) with rec...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00342-07

    authors: Fujima A,Ochiai Y,Saito A,Omori Y,Noda A,Kazuyama Y,Shoji H,Tanabayashi K,Ueda F,Yoshikawa Y,Hondo R

    更新日期:2008-01-01 00:00:00

  • Paratuberculosis infection of nonruminant wildlife in Scotland.

    abstract::Recent reports of natural paratuberculosis (or Johne's disease) in rabbits, foxes, and stoats has focused debate on the presence and importance of wildlife reservoirs in the epidemiology of this disease. This paper describes an extensive study investigating 18 nonruminant wildlife species for evidence of paratuberculo...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.39.4.1517-1521.2001

    authors: Beard PM,Daniels MJ,Henderson D,Pirie A,Rudge K,Buxton D,Rhind S,Greig A,Hutchings MR,McKendrick I,Stevenson K,Sharp JM

    更新日期:2001-04-01 00:00:00

  • Francisella tularensis Periprosthetic Joint Infections Diagnosed with Growth in Cultures.

    abstract::Tularemia caused by Francisella tularensis is a zoonotic infection of the Northern Hemisphere that mainly affects the skin, lymph nodes, bloodstream, and lungs. Other manifestations of tularemia are very rare, especially those with musculoskeletal involvement. Presenting in 2016, we diagnosed two cases of periprosthet...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章,评审

    doi:10.1128/JCM.00339-19

    authors: Chrdle A,Trnka T,Musil D,Fucentese SF,Bode P,Keller PM,Achermann Y

    更新日期:2019-07-26 00:00:00

  • Truncated human cytidylate-phosphate-deoxyguanylate-binding protein for improved nucleic acid amplification technique-based detection of bacterial species in human samples.

    abstract::A trunk of human cytidylate-phosphate-deoxyguanylate-binding protein/CXXC finger protein 1 (CFP1), immobilized onto an aminohexyl-Sepharose column, can be used as a preanalytical tool for the selective enrichment of bacterial DNA from mixed solutions with high amounts of human background DNA for nucleic acid amplifica...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02242-08

    authors: Sachse S,Straube E,Lehmann M,Bauer M,Russwurm S,Schmidt KH

    更新日期:2009-04-01 00:00:00

  • Impact of long-term storage on stability of standard DNA for nucleic acid-based methods.

    abstract::Real-time PCR is dependent upon a calibration function for quantification. While long-term storage of standards saves cost and time, solutions of DNA are prone to degradation. We present here the benchmark treatment for preservation of DNA standards, involving storage in 50% glycerol-double-distilled water, whereby a ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01230-10

    authors: Röder B,Frühwirth K,Vogl C,Wagner M,Rossmanith P

    更新日期:2010-11-01 00:00:00

  • Characterization of Micrococcaceae strains isolated from the human urogenital tract by the conventional scheme and a micromethod.

    abstract::Two hundred and twelve Micrococcaceae isolates were obtained from 82 men with nongonococcal urethritis, 24 women with vaginitis, and 54 girls with vulvovaginitis. Identification and biotyping of these strains were carried out by using the simplified scheme of Kloos and Schleifer (W. E. Kloos and K. H. Schleifer, J. Cl...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.21.3.474-477.1985

    authors: Baldellon C,Mégraud F

    更新日期:1985-03-01 00:00:00

  • COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva.

    abstract::Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of an ongoing pandemic that has infected over 36 million and killed over 1 million people. Informed implementation of government public health policies depends on accurate data on SARS-CoV-2 immunity at a population scale. We hypothesized that d...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02204-20

    authors: Pisanic N,Randad PR,Kruczynski K,Manabe YC,Thomas DL,Pekosz A,Klein SL,Betenbaugh MJ,Clarke WA,Laeyendecker O,Caturegli PP,Larman HB,Detrick B,Fairley JK,Sherman AC,Rouphael N,Edupuganti S,Granger DA,Granger SW,Coll

    更新日期:2020-12-17 00:00:00

  • Detection of immunoglobulin G to Pasteurella haemolytica capsular polysaccharide by enzyme-linked immunosorbent assay.

    abstract::An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of immunoglobulin G (IgG) to the capsular polysaccharide (CP) of Pasteurella haemolytica serotype 1. Purified CP was first covalently coupled to poly-L-lysine and then optimally adsorbed at a concentration of 5 micrograms/ml to microtiter pla...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.25.2.242-247.1987

    authors: Townsend EL,Maheswaran SK,Leininger JR,Ames TR

    更新日期:1987-02-01 00:00:00

  • Molecular relationships and antimicrobial susceptibilities of viridans group streptococci isolated from blood of neutropenic cancer patients.

    abstract::From January 1995 to May 1998, 57 episodes of bacteremia due to viridans group streptococci were identified in 50 febrile neutropenic patients with hematologic malignancies. Four patients experienced two separate episodes of streptococcal bacteremia, and one patient had four separate episodes of streptococcal bacterem...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.37.6.1876-1880.1999

    authors: Wisplinghoff H,Reinert RR,Cornely O,Seifert H

    更新日期:1999-06-01 00:00:00

  • Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by reverse transcription and polymerase chain reaction amplification.

    abstract::A rapid method for detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates, involving a combination of reverse transcription and polymerase chain reaction amplification (RT-PCR), has been developed. The RT-PCR assay employs oligonucleotide primers specific for the region of the RSV genome which enco...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.30.4.901-904.1992

    authors: Paton AW,Paton JC,Lawrence AJ,Goldwater PN,Harris RJ

    更新日期:1992-04-01 00:00:00

  • Variation in restriction fragment length polymorphisms among serial isolates from patients with Trichophyton rubrum infection.

    abstract::Molecular genotyping of strains of Trichophyton rubrum and T. mentagrophytes from patients with onychomycosis of the toes was performed to ascertain whether the fungal genotype changes over the course of time as sequential samples were obtained from patients receiving antifungal therapy and during follow-up. Sixty-six...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.39.9.3260-3266.2001

    authors: Gupta AK,Kohli Y,Summerbell RC

    更新日期:2001-09-01 00:00:00

  • Association of neurotropic viruses in HIV-infected individuals who died of secondary complications of tuberculosis, cryptococcosis, or toxoplasmosis in South India.

    abstract::The frequencies of 10 opportunistic DNA viruses were determined by multiplex real-time PCR in paired cerebrospinal fluid (CSF) and brain tissue of HIV-infected individuals. In the CSF, viruses were detectable in 45/55 cases: JC virus (JCV) in 62%, Epstein-Barr virus (EBV) in 44%, cytomegalovirus (CMV) in 25%, varicell...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.03321-12

    authors: Kannangai R,Sachithanandham J,Mahadevan A,Abraham AM,Sridharan G,Desai A,Ravi V,Shankar SK

    更新日期:2013-03-01 00:00:00

  • Rapid diagnosis of Legionella pneumophila serogroup 1 infection with the Binax enzyme immunoassay urinary antigen test.

    abstract::The Binax legionella urinary antigen (LUA) enzyme immunoassay (Binax, Portland, Maine) was evaluated in 159 patients with suspected or proven legionellosis and 209 controls. A positive LUA test was found in 37% of patients with suspected legionellosis overall and in 83% of those with proven Legionella pneumophila sero...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.35.4.954-956.1997

    authors: Kazandjian D,Chiew R,Gilbert GL

    更新日期:1997-04-01 00:00:00

  • Comparative evaluation of the API 20S and AutoMicrobic gram-positive identification systems for non-beta-hemolytic streptococci and aerococci.

    abstract::The API 20S system (Analytab Products, Plainview, N.Y.) and the AutoMicrobic Gram-Positive Identification system (GPI; Vitek Systems, Hazelwood, Mo.) were evaluated for their capacity to identify the non-beta-hemolytic streptococci and aerococci to the species level. The 20S system identified 86% (six of seven strains...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.21.4.535-541.1985

    authors: Facklam R,Bosley GS,Rhoden D,Franklin AR,Weaver N,Schulman R

    更新日期:1985-04-01 00:00:00

  • Comparison of GonoGen, GonoGen II, and MicroTrak direct fluorescent-antibody test with carbohydrate fermentation for confirmation of culture isolates of Neisseria gonorrhoeae.

    abstract::When testing 248 clinical isolates of Neisseria gonorrhoeae, the sensitivity was 100% with GonoGen (Becton Dickinson Microbiology Systems), 99.6% (247 of 248) with GonoGen II (Becton Dickinson), 97.2% (241 of 248) with the MicroTrak direct fluorescent-antibody test (Syva), and 97.6% (242 of 248) with Rapid Fermentatio...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.2.474-476.1995

    authors: Kellogg JA,Orwig LK

    更新日期:1995-02-01 00:00:00

  • Helicobacter cinaedi cellulitis and bacteremia in immunocompetent hosts after orthopedic surgery.

    abstract::At various times after orthopedic operations (more than a few weeks, with an average of 29.9 days), 11 patients had a sudden onset of high temperature (average 38.9 degrees C) and local cellulitis at different sites on the operated sides. The wounds had completely healed, without complicated infections, when the cellu...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01507-06

    authors: Kitamura T,Kawamura Y,Ohkusu K,Masaki T,Iwashita H,Sawa T,Fujii S,Okamoto T,Akaike T

    更新日期:2007-01-01 00:00:00

  • Enhanced recovery of cytomegalovirus in conventional tube cultures with a spin-amplified adsorption.

    abstract::Low-speed centrifugation-mediated adsorption was evaluated as an enhancement of infectivity of clinical and laboratory strains of cytomegalovirus (CMV) occurring with cells grown in conventional culture tubes. The time required for reporting of primary isolates of CMV from urine specimens adsorbed onto monolayers of W...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.5.965-969.1990

    authors: Oefinger PE,Shawar RM,Loo SH,Tsai LT,Arnett JK

    更新日期:1990-05-01 00:00:00

  • Latex agglutination tests for measurement of antiplague antibodies.

    abstract::A latex agglutination test was evaluated as a method for detection and titration of antiplague antibodies. Slide and microtiter techniques using polystyrene latex particles coated with specific fraction-I-antigen of Yersinia pestis were found to be comparable in specificity and sensitivity to serological tests commonl...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Suzuki S,Sakakibara H,Hotta S

    更新日期:1977-10-01 00:00:00

  • Evaluation of cell line 293 for virus isolation in routine viral diagnosis.

    abstract::Cell line 293, a continuous line of transformed human embryonic kidney cells, has been recognized for its sensitivity in the isolation of adenoviruses, particularly the fastidious species 40 and 41, from stool specimens. To explore the possibility of using this cell line for the isolation of other viruses from clinica...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.23.4.704-708.1986

    authors: Brown M,Petric M

    更新日期:1986-04-01 00:00:00