Multiplex Real-Time PCR Assay for Simultaneous Identification of Neisseria gonorrhoeae and Its Ciprofloxacin Susceptibility Status

被引：16
|
作者：
Perera, Sumudu R. [1 ,2 ]
Khan, Nurul H. [1 ,6 ]
Martin, Irene [3 ]
Taheri, Ali [1 ,7 ]
Parti, Rajinder P. [1 ,8 ]
Levett, Paul N. [4 ]
Horsman, Greg B. [4 ]
Kusalik, Anthony [5 ]
Dillon, Jo-Anne R. [1 ,2 ]
机构：
[1] Univ Saskatchewan, Vaccine & Infect Dis Org, Int Vaccine Ctr, Saskatoon, SK, Canada
[2] Univ Saskatchewan, Dept Microbiol & Immunol, Saskatoon, SK, Canada
[3] Publ Hlth Agcy Canada, Streptococcus & STI Unit, Natl Microbiol Lab, Winnipeg, MB, Canada
[4] Govt Saskatchewan, Saskatchewan Dis Control Lab, Regina, SK, Canada
[5] Univ Saskatchewan, Dept Comp Sci, Saskatoon, SK, Canada
[6] Lallemand Specialties Canada Inc, Saskatoon, SK, Canada
[7] Tennessee State Univ, Dept Agr & Environm Sci, Nashville, TN 37203 USA
[8] Evonik Transferra Nanosci, Burnaby, BC, Canada
来源：
JOURNAL OF CLINICAL MICROBIOLOGY | 2017年 / 55卷 / 11期
关键词：
Neisseria gonorrhoeae; antimicrobial susceptibility (AMS); ciprofloxacin; gyrA; multiplex RT-PCR assay; GONOCOCCAL ANTIMICROBIAL RESISTANCE; CHLAMYDIA-TRACHOMATIS; URINE SAMPLES; SURVEILLANCE; INFECTION; PERFORMANCE; PREVALENCE; MUTATIONS; GYRASE; SYSTEM;
D O I：
10.1128/JCM.00855-17
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
A real-time PCR (RT-PCR) assay was designed for the simultaneous identification of Neisseria gonorrhoeae and its ciprofloxacin susceptibility status. A SYBR green-based multiplex RT-PCR format was used; it comprised two different forward primers and a common reverse primer to detect single nucleotide polymorphisms (SNPs) in gyrA of N. gonorrhoeae. The primer pairs were evaluated for their sensitivity and specificity using genomic DNA from 254 N. gonorrhoeae isolates (82 were ciprofloxacin susceptible and 172 were ciprofloxacin resistant) and 23 non-N. gonorrhoeae species isolates. The performance of the primers was validated using genomic DNA from 100 different N. gonorrhoeae isolates (46 were ciprofloxacin susceptible and 54 were ciprofloxacin resistant) and 52 non-N. gonorrhoeae isolates. The latter panel was revalidated by testing 99 (46 isolates were ciprofloxacin susceptible and 53 isolates were ciprofloxacin resistant) of the N. gonorrhoeae isolates and 23 non-N. gonorrhoeae isolates. These primers detected N. gonorrhoeae and its ciprofloxacin susceptibility status with over 99% sensitivity and specificity for all panels tested. This assay has the potential to be an inexpensive and rapid test for the simultaneous identification of N. gonorrhoeae and its ciprofloxacin susceptibility status.
引用
收藏
页码：3201 / 3209
页数：9
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