<i>Neisseria</i>
            Species Identification Assay for the Confirmation of
            <i>Neisseria gonorrhoeae</i>
            -Positive Results of the COBAS Amplicor PCR

Mangold, Kathy A.; Regner, Mary Ann; Tajuddin, Mohammed; Tajuddin, Aamair M.; Jennings, Lawrence J.; Du, Hongyan; Kaul, Karen L.

doi:10.1128/jcm.00834-06

Cited by 28 publications

(15 citation statements)

References 27 publications

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“…When untreated, they can lead to serious consequences such as pelvic inflammatory disease, ectopic pregnancy, and infertility [4,5]. Therefore, the early and accurate diagnosis of symptomatic and asymptomatic patients is essential for preventing complications and to control the spread of infection [6]. Currently, the benefits of diagnosis and the treatment options for these infections are well understood [7].…”

Section: Introductionmentioning

confidence: 99%

Performance of the Abbott Real Time CT/NG assay in urines and cervico-vaginal samples from Senegal

Gueye

Diop-Ndiaye

Gningue

et al. 2014

J Infect Dev Ctries

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Introduction: Chlamydia trachomatis and Neisseria gonorrhoeae are the most common causes of sexually transmitted disease in Senegal and worldwide. Molecular techniques have become the standard for their detection, and due to the frequency of co-infections, these tests can detect both agents and can be used on urine samples, vaginal swabs, or endocervical samples. In developing countries, the use of these molecular techniques is very limited and there is a need for evaluations of these techniques to be done. Methodology: A total of 181 samples were tested with the Abbott RealTime CT/NG assay and compared with the Roche Cobas Amplicor CT/NG assay. Specimens were collected from the key population of men having sex with men (urine, n = 60), female sex workers (genital swabs, n = 60) and from women visiting the laboratory for a gynecological checkup (urine, n = 60 and endocervical samples, n = 61). Results: The agreement between the two techniques was 98.90% with a Kappa coefficient of 0.98. A sensitivity of 93.3%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 93.3% were found for both Chlamydia trachomatis and Neisseria gonorrhoeae. Conclusion: These results showed that both methods are similar and suitable for the detection of CT/NG in all types of samples examined in this study.

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Section: Introductionmentioning

confidence: 99%

Performance of the Abbott Real Time CT/NG assay in urines and cervico-vaginal samples from Senegal

Gueye

Diop-Ndiaye

Gningue

et al. 2014

J Infect Dev Ctries

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“…The real-time curve was, however, morphologically similar to those that amplified at earlier cycles. The NAAT discordance may reflect a false positive on opa gene testing or a false negative from porA pseudogene testing, which has been found by some authors to be less sensitive than other confirmatory NAATs (6). A headto-head comparison of the opa gene on the m2000 and the porA pseudogene method PCR found that the m2000 had a 10-fold greater analytical sensitivity (T. Anderson and M. J. Maze, unpublished data).…”

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confidence: 99%

“…All but one specimen with DNA detected that was unsupported by culture had DNA detected by the use of the porA pseudogene. The porA pseudogene has been validated previously as a suitable confirmatory test for positive NAATs (6,8,9). The combination of an appropriate clinical history and positive confirmatory NAAT makes a false-positive result very unlikely.…”

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confidence: 99%

Nucleic Acid Amplification of the opa Gene for Detection of Neisseria gonorrhoeae: Experience from a Diagnostic Laboratory

et al. 2011

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We report results of Neisseria gonorrhoeae nucleic acid amplification testing (NAAT) with the Abbott m2000 PCR at a tertiary laboratory 6 months after its introduction. Of 5,475 specimens tested, 45 samples (0.82%) tested positive for N. gonorrhoeae. Eight were not cultured, but seven tested positive with a porA pseudogene NAAT.Neisseria gonorrhoeae is a sexually transmitted bacterium that causes a spectrum of disease ranging from asymptomatic infection to pelvic inflammatory disease. Gonorrhea is not a notifiable disease in New Zealand, and therefore accurate estimation of the prevalence is difficult. The prevalence has been estimated at 90 cases/100,000 population (2), although this is likely to be an underestimation as it includes only those cases diagnosed at sexual health and family planning clinics. The presumed rate is about 6 times lower than that of chlamydia infection.Nucleic acid amplification testing (NAAT) was introduced at Canterbury Health Laboratories in addition to culture due to faster turnaround-time (7). Testing is routinely performed with the Abbott m2000 as a duplex PCR for Chlamydia trachomatis and N. gonorrhoeae on all specimens with requests for detection of either microorganism. Since introduction, all specimens with a positive NAAT result have undergone testing with a confirmatory NAAT of the porA pseudogene if they have not been cultured. Routine testing with the Abbott m2000 is performed using Abbott multi-Collect specimen collection kits (MML Diagnostics Packaging, Inc., Troutdale, OR) according to the manufacturer's instructions. The porA pseudogene testing was performed by the method described by Wiley et al. (9). Culture was performed on Thayer-Martin agar (Fort Richard Laboratories, Ltd., New Zealand) in 5% carbon dioxide for 3 days.Most reports in the literature report results from NAATs of sample collections with a high proportion containing Neisseria gonorrhoeae (5, 7). In contrast, reports from diagnostic use of earlier generation NAATs found low positive predictive values and false positives. Concerns have been raised about the high rates of false-positive results in populations with a low prevalence of N. gonorrhoeae (1,3,4,10).New Zealand currently conducts NAAT screening for chlamydia in sexually active populations. Addition of N. gonorrhoeae by NAAT means that potentially low-risk population groups would be tested and false-positive results in settings with a low positive predictive value could be detrimental to the screened population.The objective of this retrospective review was to review cases of N. gonorrhoeae that were diagnosed solely by NAAT and assess the occurrence of likely false-positive results.All specimens tested by PCR for N. gonorrhoeae between 1 November 2009 and 1 May 2010 were retrospectively reviewed. The study was determined to be exempt from review by the regional ethics committee. Those specimens that had a positive NAAT had the results of their culture or confirmatory NAAT reviewed. The information was analyzed to determine the proportion of sp...

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confidence: 99%

“…DNA probes, such as those used in real-time PCR assays, are able to bind to only a limited range of DNA sequences. Thus, currently available PCR-based detection methods are usually limited to differentiating among bacterial species within a single genus (8,11,20,26). Bacterial species identification can be moderately increased by multiplexing several probes to perform different species detection assays simultaneously (18,30); however, the fundamental limitation caused by a narrow probe binding range remains the same.…”

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confidence: 99%

Rapid Universal Identification of Bacterial Pathogens from Clinical Cultures by Using a Novel Sloppy Molecular Beacon Melting Temperature Signature Technique

Chakravorty

Aladegbami

Burday³

et al. 2010

J Clin Microbiol

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A real-time PCR assay with the ability to rapidly identify all pathogenic bacteria would have widespread medical utility. Current real-time PCR technologies cannot accomplish this task due to severe limitations in multiplexing ability. To this end, we developed a new assay system which supports very high degrees of multiplexing. We developed a new class of mismatch-tolerant "sloppy" molecular beacons, modified them to provide an extended hybridization range, and developed a multiprobe, multimelting temperature (T m ) signature approach to bacterial species identification. Sloppy molecular beacons were exceptionally versatile, and they were able to generate specific T m values for DNA sequences that differed by as little as one nucleotide to as many as 23 polymorphisms. Combining the T m values generated by several probe-target hybrids resulted in T m signatures that served as highly accurate sequence identifiers. Using this method, PCR assays with as few as six sloppy molecular beacons targeting bacterial 16S rRNA gene segments could reproducibly classify 119 different sequence types of pathogenic and commensal bacteria, representing 64 genera, into 111 T m signature types. Blinded studies using the assay to identify the bacteria present in 270 patient-derived clinical cultures including 106 patient blood cultures showed a 95 to 97% concordance with conventional methods. Importantly, no bacteria were misidentified; rather, the few species that could not be identified were classified as "indeterminate," resulting in an assay specificity of 100%. This approach enables highly multiplexed target detection using a simple PCR format that can transform infectious disease diagnostics and improve patient outcomes.Human bloodstream infections (BSI) must be treated rapidly and effectively in order to avoid significant morbidity and mortality (17). A rising incidence of drug-resistant infections has complicated antibiotic selection (33), emphasizing the importance of rapidly determining the identity and drug susceptibility profile of each infecting bacterial species. Unfortunately, conventional microbiological identification and drug susceptibility determination methods are often too time-consuming to allow quick treatment decisions. Many bacterial species have specific antibiotic indications while most have local antibiotic resistance patterns that can be predicted by periodic examination of antibiotic resistance profiles (10,19). Consequently, species identification may be used to guide antibiotic therapy pending final antibiotic susceptibility tests. A rapid bacterial species identification method would dramatically speed up the diagnosis of serious diseases, enabling rapid and definitive treatment, and concomitantly decrease the use of broad-spectrum antibiotics.Ideally, a rapid assay for BSI should be functionally equivalent to the current diagnostic standard, blood culture (15), in being able to identify all clinically significant pathogens as well as commensals. To this end, molecular assays have targeted bacterial 1...

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Neisseria Species Identification Assay for the Confirmation of Neisseria gonorrhoeae -Positive Results of the COBAS Amplicor PCR

Cited by 28 publications

References 27 publications

Performance of the Abbott Real Time CT/NG assay in urines and cervico-vaginal samples from Senegal

Performance of the Abbott Real Time CT/NG assay in urines and cervico-vaginal samples from Senegal

Nucleic Acid Amplification of the opa Gene for Detection of Neisseria gonorrhoeae: Experience from a Diagnostic Laboratory

Rapid Universal Identification of Bacterial Pathogens from Clinical Cultures by Using a Novel Sloppy Molecular Beacon Melting Temperature Signature Technique

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