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Development of a SimpleProbe real-Time PCR Assay for rapid detection and identification of the US novel urethrotropic clade of Neisseria meningitidis ST-11 (US_NmUC).
- Source :
-
PloS one [PLoS One] 2020 Feb 10; Vol. 15 (2), pp. e0228467. Date of Electronic Publication: 2020 Feb 10 (Print Publication: 2020). - Publication Year :
- 2020
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Abstract
- Urethritis, or inflammation of the urethra, is one of the most common reasons men seek clinical care. Sexually transmitted pathogens including Neisseria gonorrhoeae are responsible for over half of the symptomatic urethritis cases in U.S. men. Recently, clinics in Indianapolis, Columbus, Atlanta, and other U.S. cities began to note increasing numbers of men presenting with urethritis and Gram-negative intracellular diplococci in their urethral smears who test negative for N. gonorrhoeae. Many of these discordant cases, which have periodically reached highs of more than 25% of presumed gonococcal cases in some sexually transmitted infection clinics in the U.S. Midwest, are infected with strains in a novel urethrotropic clade of Neisseria meningitidis ST-11 (US&#95;NmUC). However, no cultivation-independent tests are available for the US&#95;NmUC strains, and prior studies relied on microbial culture and genome sequencing to identify them. Here, we describe a PCR test that can identify the US&#95;NmUC strains and distinguish them from commensal and invasive N. meningitidis strains as well as N. gonorrhoeae. Our SimpleProbe®-based real-time PCR assay targets a conserved nucleotide substitution in a horizontally acquired region of US&#95;NmUC strain genomes. We applied the assay to 241 urine specimens whose microbial compositions had previously been determined by deep shotgun metagenomic sequencing. The assay detected the single US&#95;NmUC positive case in this cohort, with no false positives. Overall, our simple and readily adaptable assay could facilitate investigation of the pathogenesis and epidemiology of the US&#95;NmUC clade.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Adolescent
Adult
Aged
Aged, 80 and over
Case-Control Studies
False Positive Reactions
Gonorrhea diagnosis
Gonorrhea epidemiology
Gonorrhea microbiology
Gonorrhea urine
Humans
Male
Middle Aged
Neisseria gonorrhoeae genetics
Neisseria gonorrhoeae isolation & purification
Neisseria meningitidis classification
Neisseria meningitidis isolation & purification
Polymorphism, Single Nucleotide
Sexually Transmitted Diseases, Bacterial diagnosis
Sexually Transmitted Diseases, Bacterial epidemiology
Sexually Transmitted Diseases, Bacterial microbiology
Sexually Transmitted Diseases, Bacterial urine
United States epidemiology
Urethra microbiology
Urethra pathology
Urethritis diagnosis
Urinalysis methods
Whole Genome Sequencing
Young Adult
Neisseria meningitidis genetics
Real-Time Polymerase Chain Reaction methods
Urethritis microbiology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 15
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 32040516
- Full Text :
- https://doi.org/10.1371/journal.pone.0228467