Detection Of Neisseria Gonorrhoeae Research Articles

Performance evaluation of loop-mediated isothermal amplification, polymerase chain reaction and real-time polymerase chain reaction methods to detect Neisseria gonorrhoeae among symptomatic patients from India.

Background: Neisseria gonorrhoeae is one of the most important causative organisms in causing sexually transmitted infections. The clinical presentation of gonorrhoea mimics the symptoms of other sexually transmitted infections, and a proper diagnosis of the same is therefore crucial in patient management. The current study intended to compare different in-house molecular methods: that is, conventional PCR, real-time PCR, and LAMP assay for detection of N. gonorrhoeae. Methods: A total of 163 samples were collected from 145 patients who presented with urethral and vaginal discharge. Collected samples were processed for culture on GC agar base, and three different molecular diagnostic tests (conventional PCR, real-time PCR, and LAMP assay) were performed simultaneously on all the samples. Results: Culture of N. gonorrhoeae was positive in 17 out of 21 (80.9%) swab samples. With culture as the gold standard method, conventional and real-time PCR had a sensitivity of 94.1%, whereas the sensitivity of the LAMP assay was found to be 88.2%. All three methods had a specificity of 100%. In addition to swab samples, evaluation of urine samples by different molecular methods yielded a good concordance with a kappa value of 0.85 by conventional PCR and real-time PCR showing a perfect level of agreement, while the LAMP assay was found to have a substantial level of agreement. Conclusion: LAMP assay had a comparable diagnostic accuracy to other molecular methods for the detection of N. gonorrhoeae and can be used as a point-of-care test in resource-limited settings.

Self-sampling with oral rinse to detect oropharyngeal Neisseria gonorrhoeae among men who have sex with men: results from an exploratory study in Belgium (the SSONG Study)

ObjectivesWe aimed to assess whether a self-collected oral rinse was non-inferior to clinician-collected oropharyngeal swabs to detect Neisseria gonorrhoeae (Ng) using culture and nucleic acid amplification tests (NAAT) among men...

Neisseria gonorrhoeae prevalence in females in São Paulo, Brazil: surveillance of the infection over a 11-year period.

The prevalence of infection by Neisseria gonorrhoeae is the second highest among bacterial sexually transmitted infections worldwide. It can lead to severe complications, especially in the female reproductive system. So, the objective of the present study was to assess the prevalence of Neisseria gonorrhoeae infection in a large population of female patients from a private health service in São Paulo (Brazil), thereby identifying the main age groups affected by the agent and how prevalence progressed over time. A cross-sectional study was conducted based on the results of all molecular biology tests performed for detecting Neisseria gonorrhoeae. The tests were conducted between January 2005 and December 2015. Positive test results were grouped by year and age group. Of those tests, 35,886 were considered eligible for the statistical data. The overall prevalence of Neisseria gonorrhoeae infection in the study population was 0.4%. A higher prevalence of infection was observed in the group whose participants' age was ≤ 25 years, at a rate of 0.6%. There was no significant increase or decrease in the number of positive test results over time. The prevalence of the infection in those aged between 10 and 19 years, 20-29 years, 30-39 years, 40-49 years, 50-59 years, and 60 years or above was respectively 0.87%, 0.50%, 0.36%, 0.22%, 0.17%, and 0.26%. The screening of asymptomatic young women would have the potential to reduce infection, transmission, and sequelae of infection by this agent.

Usability of a novel lateral flow assay for the point-of-care detection of Neisseria gonorrhoeae: A qualitative time-series assessment among healthcare workers in South Africa.

Accurate and user-friendly rapid point-of-care diagnostic tests (POCT) are needed to optimize treatment of Neisseria gonorrhoeae, especially in low-resource settings where syndromic management is the standard of care for sexually transmitted infections. This study aimed to assess the acceptability and usability of a novel lateral flow assay and portable reader for the point-of-care detection of N. gonorrhoeae infection (NG-LFA). This mixed-methods study was conducted as part of a diagnostic performance and usability evaluation of a prototype NG-LFA for detection of N. gonorrhoeae in symptomatic men and women at primary healthcare facilities in the Buffalo City Metro, South Africa. The Standardized System Usability Scale (SUS) was administered, and in-depth interviews were conducted among healthcare professionals (HCPs) and fieldworkers (FWs) at pre-implementation, initial use and 3- and 6-month study implementation to assess user expectations, practical experience, and future implementation considerations for the NG-LFA. Data collection and analysis was guided by the Health Technology Adoption Framework, including new health technology attributes, learnability, satisfaction, and suitability. The framework was adapted to include perceived durability. A total of 21 HCPs and FWs were trained on the NG-LFA use. SUS scores showed good to excellent acceptability ranging from 78.8-90.6 mean scores between HCPs and FWs across study time points. All transcripts were coded using Dedoose and qualitative findings were organized by learnability, satisfaction, suitability, and durability domains. Usability themes are described for each time point. Initial insecurity dissipated and specimen processing dexterity with novel POCT technology was perfected over time especially amongst FWs through practical learning and easy-to-use instructions (learnability). Participants experienced both positive and negative test results, yielding perceived accuracy and minimal testing challenges overall (satisfaction). By 3- and 6-month use, both HCPs and FWs found the NG-LFA convenient to use in primary health care facilities often faced with space constraints and outlined perceived benefits for patients (suitability and durability). Findings show that the NG-LFA device is acceptable and usable even amongst paraprofessionals. High SUS scores and qualitative findings demonstrate high learnability, ease-of-use and suitability that provide valuable information for first-step scale-up requirements at primary healthcare level. Minor prototype adjustments would enhance robustness and durability aspects.

A new real-time quantitative polymerase chain reaction method using locked nucleic acids to detect Neisseria gonorrhoeae infection and point mutation on gyrA associated with quinolone susceptibility

In Neisseria gonorrhoeae, fluoroquinolone resistance is caused by mutations on various genes, especially on gyrA. Locked nucleic acid DNA sequence was added to the forward primer for gyrA and subjected to real-time quantitative polymerase chain reaction, enabling simultaneous detection of N. gonorrhoeae and mutations associated with antimicrobial resistance.

Proof-of-concept, rapid, instrument-free molecular detection of Neisseria gonorrhoeae and ciprofloxacin susceptibility

To develop instrument-free point-of-care methods using recombinase polymerase amplification (RPA) technology coupled with a simple lateral flow detection system to detect Neisseria gonorrhoeae and susceptibility to ciprofloxacin. For identification of gonococcal infection, an RPA-based method was developed targeting the gonococcal porA pseudogene (NG-porA-RPA). For ciprofloxacin susceptibility, predictive WT sequences at codons 91 and 95 of the gonococcal gyrA DNase gene were targeted. Given the known complexities of SNP detection using RPA (e.g. the ability to accommodate mismatches) we trialled several different assays incorporating various additional non-template mismatches in the oligonucleotide sequences to reduce affinity for the mutant (resistant) gyrA sequences. Assays were evaluated using a bank of N. gonorrhoeae (n = 10) and non-gonococcal (n = 5) isolates and a panel of N. gonorrhoeae nucleic acid amplification test (NAAT)-positive clinical sample extracts (n = 40). The NG-porA-RPA assay was specific to N. gonorrhoeae and provided a positive percentage agreement (PPA) of 87.5% (35/40) compared with a commercial N. gonorrhoeae NAAT when applied to the 40 clinical sample extracts. For gyrA, the non-template bases successfully reduced banding intensity for double-mutant strains (mutations at both 91 and 95), but not for rarer single-mutant (91 only) strains. The most promising gyrA assay, NG-gyrA-RPA08, correctly detected 83% (25/30) of infections from NAAT-positive clinical samples confirmed to have WT gyrA sequences based on Sanger sequencing. These proof-of-concept data show that RPA technology has considerable promise for detecting N. gonorrhoeae and associated antibiotic susceptibility and would offer a diagnostic-based stewardship strategy identified as urgently needed by the WHO.

Prevalence of nonviral reproductive tract infections/sexually transmitted infections in female patients with cervicovaginal discharge: Excerpts from a regional reference center in North India.

To study the prevalence of common nonviral reproductive tract infections/sexually transmitted infections (RTI/STI) prevailing among females who presented to our regional STI reference center and to ascertain the association of various symptoms with different RTI/STIs. A retrospective analysis of female patients presenting to our STI Regional center located in the Department of Medical Microbiology in PGIMER, Chandigarh, was done between April 2018 and December 2019 for patients presenting with cervico-vaginal discharge. Two to three swabs were collected from each patient. The first swab was subjected to wet mount, gram stain, Potassium hydroxide (KOH) test, and culture on blood agar, the colonies obtained were identified by matrix-assisted laser desorption time of flight mass spectrometer (MALDI TOF-MS). Second swab was used for DNA extraction and detection of Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV), and Chlamydia trachomatis (CT) by polymerase chain reaction. The third swab, when available, was inoculated onto pleuropneumonia-like organisms (PPLO) broth. One thousand and thirteenth of 1472 (69%) female patients were symptomatic and the most common presenting symptoms were vaginal discharge (707/1013 [69.8%]), infertility (266/1013 [26.2%]), genital itching (60/1013 [5.9%]), lower abdomen pain (47/1013 [4.6%]) and burning micturition (16/1013 [1.6%]). The most prevalent RTI/STI was bacterial vaginosis (BV) 18.2% (269/1472), followed by vulvovaginal candidiasis (VVC) 6.8% (100/1472) and trichomoniasis (TV) 1.9% (28/1472). Five cases each of Mycoplasma genitalium and Ureaplasma urealyticum, three of NG and one of CT were also identified. Coinfections were seen in 40 (2.7%) cases. The most common causative agent responsible for VVC in our study was Candida albicans (65%). RTI/STIs were common among women and 69% were symptomatic. BV was the most common STI present in 18.2%, followed by VVC (6.8%) and trichomoniasis (1.9%).

A statistical review: why average weighted accuracy, not accuracy or AUC?

Sensitivity and specificity are key aspects in evaluating the performance of diagnostic tests. Accuracy and AUC are commonly used composite measures that incorporate sensitivity and specificity. Average Weighted Accuracy (AWA) is motivated by the need for a statistical measure that compares diagnostic tests from the medical costs and clinical impact point of view, while incorporating the relevant prevalence range of the disease as well as the relative importance of false-positive versus false-negative cases. We illustrate the testing procedures in four different scenarios: (i) one diagnostic test vs. the best random test, (ii) two diagnostic tests from two independent samples, (iii) two diagnostic tests from the same sample, and (iv) more than two diagnostic tests from different or the same samples. The impacts of sample size, prevalence, and relative importance on power and average medical costs/clinical loss are examined through simulation studies. Accuracy has the highest power while AWA provides a consistent criterion in selecting the optimal threshold and better diagnostic tests with direct clinical interpretations. The use of AWA is illustrated on a three-arm clinical trial evaluating three different assays in detecting Neisseria gonorrhoeae and Chlamydia trachomatis in the rectum and pharynx.

MULTIPLEX POLYMERASE CHAIN REACTION (PCR) FOR THE DETECTION OF NEISSERIA GONORRHOEAE AND THE QUINOLONE RESISTANCE GENE IN PAKISTAN

Objective: To detect Neisseria gonorrhoeae from the urine of male patients reporting with active urethral discharge using multiplex polymerase chain reaction (PCR). And the simultaneous detection of the quinolone resistance determining region (QRDR) on the Neisseria gonorrhoeae gene using multiplex polymerase chain reaction.
 Study Design: Cross sectional study.
 Place and Duration of Study: Microbiology department, Army Medical College Rawalpindi Pakistan, from Mar to Dec 2018.
 Methodology: Male patients with active urethral discharge with no past history of antibiotic use for urethral discharge were included in study and patients without active urethral discharge and history of antibiotic use for urethral discharge were excluded. Urine of patients of active urethral discharge was collected and multiplex polymerase chain reaction was done by using two forward primers along with common reverse primer.
 Results: In this study 24 (40%) of patients who presented with active urethral discharge were positive for gonorrhea. However Quinolone Resistance Determining Region is detected in 17 (70.83%) of cases and only 7 (29.17%) were sensitive to ciprofloxacin.
 Conclusion: The multiplex polymerase chain reaction is very efficient and effective method for the simultaneous detection of Neisseria gonorrhoeae and status of isolate susceptibility to ciprofloxacin. And in Pakistan ciprofloxacin cannot be used as first line drug for the treatment of gonorrhea.

Detection of Neisseria gonorrhoeae and Chlamydia trachomatis infections in pregnant women by multiplex recombinase polymerase amplification.

Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are the main pathogenic microorganisms causing sexually transmitted infections. In this study, a multiplex thermostable recombinase polymerase amplification-lateral flow detection (RPA-LFD) assay was established, and the reaction conditions such as the ratio of primer concentration, magnesium ion concentration, amplification time and template DNA concentration in the multiplex RPA reaction were optimized. The optimized multiplex RPA-LFD method was used to detect both CT and NG positive control plasmids, and it was found that the LFD could be used to obtain visible results when the plasmid copy number was only 200. The sensitivity of the multiplex RPA-LFD method used for clinical samples was 85.62 (95% CI at 53.66–97.29) for NG detection and 90.90 (95% CI at 57.12–99.52) for CT detection.

Using rapid point-of-care tests to inform antibiotic choice to mitigate drug resistance in gonorrhoea.

BackgroundThe first cases of extensively drug resistant gonorrhoea were recorded in the United Kingdom in 2018. There is a public health need for strategies on how to deploy existing and novel antibiotics to minimise the risk of resistance development. As rapid point-of-care tests (POCTs) to predict susceptibility are coming to clinical use, coupling the introduction of an antibiotic with diagnostics that can slow resistance emergence may offer a novel paradigm for maximising antibiotic benefits. Gepotidacin is a novel antibiotic with known resistance and resistance-predisposing mutations. In particular, a mutation that confers resistance to ciprofloxacin acts as the ‘stepping-stone’ mutation to gepotidacin resistance.AimTo investigate how POCTs detecting Neisseria gonorrhoeae resistance mutations for ciprofloxacin and gepotidacin can be used to minimise the risk of resistance development to gepotidacin.MethodsWe use individual-based stochastic simulations to formally investigate the aim.ResultsThe level of testing needed to reduce the risk of resistance development depends on the mutation rate under treatment and the prevalence of stepping-stone mutations. A POCT is most effective if the mutation rate under antibiotic treatment is no more than two orders of magnitude above the mutation rate without treatment and the prevalence of stepping-stone mutations is 1–13%.ConclusionMutation frequencies and rates should be considered when estimating the POCT usage required to reduce the risk of resistance development in a given population. Molecular POCTs for resistance mutations and stepping-stone mutations to resistance are likely to become important tools in antibiotic stewardship.

Detection of Neisseria gonorrhoeae from Joint Aspirate by Metagenomic Sequencing in Disseminated Gonococcal Infection.

Disseminated gonococcal infection (DGI) often manifests as gonococcal arthritis and may carry significant morbidity. However, diagnosis remains elusive due to limited sensitivity of available diagnostic tests. We used metagenomic next-generation sequencing to detect Neisseria gonorrhoeae from culture-negative joint aspirates of 2 patients with clinically diagnosed DGI.

Developing target product profiles for Neisseria gonorrhoeae diagnostics in the context of antimicrobial resistance: An expert consensus.

BackgroundThere is a need for a rapid diagnostic point of care test to detect Neisseria gonorrhoeae (NG) infection to prevent incorrect, lack or excess of treatment resulting from current syndromic management in low-resource settings. An assay to identify NG antimicrobial resistance (AMR) is also highly desirable to facilitate antibiotic stewardship. Here we describe the development of two target product profiles (TPPs): one for a test for etiological diagnosis of NG and Chlamydia trachomatis (CT) (TPP1) and one for the detection of NG AMR/susceptibility (TPP2).MethodsDraft TPPs were initially developed based on a landscape analysis of existing diagnostics and expert input. TPPs were refined via an online Delphi survey with two rounds of input from 68 respondents. TPP characteristics on which <75% of non-industry respondents agreed were further discussed and revised by an expert working group.ResultsThe need for a test to identify NG in patients with urethral or vaginal discharge was identified as a minimal requirement of TPP1, with a test that can diagnose NG in asymptomatic patients as the optimal requirement. A sensitivity of 80% was considered acceptable, either in context of syndromic management or screening high-risk populations. For TPP2, the agreed minimal requirement was for a test to be used at level 2 healthcare facilities and above, with an optimal requirement of level 1 or above. A lateral flow format was preferred for TPP1, while it was considered likely that TPP2 would require a molecular format. A total of 31 test characteristics were included in TPP1 and 27 in TPP2.ConclusionsFollowing the working group revisions, TPPs were posted online for public feedback for two months, and are now finalized. The final TPPs are currently guiding the development of new diagnostics that meet the defined characteristics to reach the market within two years.

Diag nostics of gonorrhoea in Ukraine following World Health Organization recommendations

Objective — to evaluate the compliance of gonorrhoea diagnostics in regional skin and venereological dispensaries (RSVD) in Ukraine during 2013—2014 to World Health Organization (WHO) laboratory recommendations and identify areas for its improvement. Materials and methods. The study analyzed statistics data regarding the number of performed laboratory tests and their positive results for the detection of gonococci, the collection of biological samples and the use of laboratory methods for N. gonorrhoeae identification obtained from 14 RSVD of Ukraine by questionnaire. Results and discussion. Dispensary laboratories performed microscopy, culture and point-of-care-tests in 100, 85.7 and 7.1 % of patients, respectively. Gonococci were found in 2.3 % of the patients, equally by the results of microscopy (1740/76 348) and by culture (1322/56 273). Dispensary laboratories did not perform nucleic acid amplification tests. Sampling of biological material from three anatomical sites (urogenital, pharyngeal and rectal specimens) in heterosexual patients with symptoms of sexually transmitted infection was performed by 25 % of respondents, from two sites (urogenital and pharyngeal) — by 41.7 %, and from one site (urogenital sample) — by 33.3 % of dispensaries. External quality control of laboratory tests for the detection of N. gonorrhoeae was performed by 50 % of the respondents. Non-selective nutrient media for isolation of N. gonorrhoeae was used by 16.7 % of dispensaries, and prepared nutrient media in test tubes instead of the recommended Petri dishes, were used in 58.3 % of dispensaries. The vast majority of respondents (78.6 %) had an experience of determining gonococcal sensitivity to antibiotics. Two institutions (14.3 %) participated in an international pilot project to determine the antibiotic resistance of gonococci. Conclusions. Diagnostics of gonococcal infection in Ukraine is not optimal and needs improvement by using quality-tested bacteriological cultures and point-of-care-tests, implementation of nucleic acid amplification tests, monitoring of gonococcal sensitivity to antibiotics, collection of extragenital biological samples. Adherence to evidence-based international guidelines and adapted national guidelines for the diagnostics of gonorrhoea is an essential factor of the controlling of gonococcal infection.

The Laboratory Diagnosis of Neisseria gonorrhoeae: Current Testing and Future Demands.

The ideal laboratory test to detect Neisseria gonorrhoeae (Ng) should be sensitive, specific, easy to use, rapid, and affordable and should provide information about susceptibility to antimicrobial drugs. Currently, such a test is not available and presumably will not be in the near future. Thus, diagnosis of gonococcal infections presently includes application of different techniques to address these requirements. Microscopy may produce rapid results but lacks sensitivity in many cases (except symptomatic urogenital infections in males). Highest sensitivity to detect Ng was shown for nucleic acid amplification technologies (NAATs), which, however, are less specific than culture. In addition, comprehensive analysis of antibiotic resistance is accomplished only by in vitro antimicrobial susceptibility testing of cultured isolates. As a light at the end of the tunnel, new developments of molecular techniques and microfluidic systems represent promising opportunities to design point-of-care tests for rapid detection of Ng with high sensitivity and specificity, and there is reason to hope that such tests may also provide antimicrobial resistance data in the future.

Clinical Evaluation of the Click Sexual Health Test for the Detection of Neisseria Gonorrhoeae (NG), Trichomonas Vaginalis (TV), and Chlamydia Trachomatis (CT) in Women

Clinical Evaluation of the Click Sexual Health Test for the Detection of Neisseria Gonorrhoeae (NG), Trichomonas Vaginalis (TV), and Chlamydia Trachomatis (CT) in Women

Comparison of an in-house real-time duplex PCR assay with commercial HOLOGIC\xae APTIMA assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis in urine and extra-genital specimens

BackgroundExtra-genital Neisseria gonorrhoeae and Chlamydia trachomatis infections are mostly asymptomatic, and important reservoir sites of infection as they often go undetected and may be more difficult to eradicate with recommended therapeutic regimens. Commercial nucleic acid amplification tests (NAATs) have not received regulatory approval for the detection of N. gonorrhoeae and C. trachomatis in extra-genital specimens. The HOLOGIC® APTIMA Combo2 assay for N. gonorrhoeae and C. trachomatis has performed well in evaluations using extra-genital specimens.MethodsWe assessed the performance of an in-house real-time duplex PCR assay for the detection of N. gonorrhoeae and C. trachomatis in urine and extra-genital specimens using the HOLOGIC® APTIMA assays as gold standard comparators. Urine, oropharyngeal and ano-rectal specimens were collected from each of 200 men-who-have-sex-with-men (MSM) between December 2011 and July 2012.ResultsFor N. gonorrhoeae detection, the in-house PCR assay showed 98.5–100% correlation agreement with the APTIMA assays, depending on specimen type. Sensitivity for N. gonorrhoeae detection was 82.4% for ano-rectal specimens, 83.3% for oropharyngeal specimens, and 85.7% for urine; and specificity was 100% with all specimen types. The positive predictive value (PPV) for N. gonorrhoeae detection was 100% and the negative predictive value (NPV) varied with sample type, ranging from 98.5–99.5%. For C. trachomatis detection, correlation between the assays was 100% for all specimen types. The sensitivity, specificity, PPV and NPV of the in-house PCR assay was 100% for C. trachomatis detection, irrespective of specimen type.ConclusionThe in-house duplex real-time PCR assay showed acceptable performance characteristics in comparison with the APTIMA® assays for the detection of extra-genital N. gonorrhoeae and C. trachomatis.

Antimicrobial Resistance in Neisseria gonorrhoeae and Treatment of Gonorrhea.

Gonorrhea and antimicrobial resistance (AMR) in Neisseria gonorrhoeae are major public health concerns globally. Dual antimicrobial therapy (mainly ceftriaxone 250-500mg×1 plus azithromycin 1-2g×1) is currently recommended in many countries. These dual therapies have high cure rates, have likely been involved in decreasing the level of cephalosporin resistance internationally, and inhibit the spread of AMR gonococcal strains. However, ceftriaxone-resistant strains are currently spreading internationally, predominately associated with travel to Asia. Furthermore, the first global treatment failure with recommended dual therapy was reported in 2016 and the first isolates with combined ceftriaxone resistance and high-level azithromycin resistance were reported in 2018 in the UK and Australia. New antimicrobials for treatment of gonorrhea are essential and, of the few antimicrobials in clinical development, zoliflodacin particularly appears promising. Holistic actions are imperative. These include an enhanced advocacy; prevention, early diagnosis, contact tracing, treatment, test-of-cure, and additional measures for effective management of anogenital and pharyngeal gonorrhea; antimicrobial stewardship; surveillance of infection, AMR and treatment failures; and intensified research, for example, regarding rapid molecular point-of-care detection of gonococci and AMR, novel AMR determinants, new antimicrobials, and an effective gonococcal vaccine, which is the only sustainable solution for management and control of gonorrhea.

Clinical evaluation of dry chemical enzyme method for detecting Neisseria gonorrhoeae and analysis of external quality assessment results

Objective:Neisseria gonorrhoeae in female cervix sample was detected by dry chemical enzymatic method and culture method. The detection effects of two detection methods were analyzed to provide reference for clinical detection. At the same time, strains were separated and identified to evaluate reliability of different methods. Methods: During October 2015 to December 2017, 8 860 samples of female cervix from the Third Affiliated Hospital of Zhengzhou University and Henan Provincial Peoples Hospital were detected by dry chemical enzymatic method and culture method. Because of the possible leak detection by culture method, the inconsistent results were supplemented by real-time PCR assay. The data were analyzed by SPSS 19.0. Fifteen strains of external quality assessment of Neisseria gonorrhoeae identification and validation during 2015 to 2017 years were identified by latex chromatography, culture method, dry chemical enzyme method and PCR- fluorescent probe method. Results: The positive rates of dry chemical enzymatic method and culture method for detection of Neisseria gonorrhoeae were 0.88% (78/8 860) and 0.41% (36/8 860), respectively. Among these, 26 samples were tested positive by dry chemical enzymatic method which were consistent with real-time PCR assay. Meanwhile, the culture method was negative. Dry chemistry enzyme method external quality assessment results of 3 years were accurate. Conclusion: The positive rate of dry chemical enzymatic method was markedly higher than that of culture method (P<0.05). The dry chemical enzymatic method, while exhibiting a high specificity, had high detection rate compared to culture method. And external quality assessment results are accurate. The clinical coincidence rate of dry chemical enzymatic detection was higher.

A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples.

Globally, the microbe Neisseria gonorrhoeae (NG) causes 106 million newly documented sexually transmitted infections each year. Once appropriately diagnosed, NG infections can be readily treated with antibiotics, but high-risk patients often do not return to the clinic for treatment if results are not provided at the point of care. A rapid, sensitive molecular diagnostic would help increase NG treatment and reduce the prevalence of this sexually transmitted disease. Here, we report on the design and development of a rapid, highly sensitive, paperfluidic device for point-of-care diagnosis of NG. The device integrates patient swab sample lysis, nucleic acid extraction, thermophilic helicase-dependent amplification (tHDA), an internal amplification control (NGIC), and visual lateral flow detection within an 80min run time. Limits of NG detection for the NG/NGIC multiplex tHDA assay were determined within the device, and clinical performance was validated retroactively against qPCR-quantified patient samples in a proof-of-concept study. This paperfluidic diagnostic has a clinically relevant limit of detection of 500 NG cells per device with analytical sensitivity down to 10 NG cells per device. In triplicate testing of 40 total urethral and vaginal swab samples, the device had 95% overall sensitivity and 100% specificity, approaching current laboratory-based molecular NG diagnostics. This diagnostic platform could increase access to accurate NG diagnoses to those most in need.