Implementation and evaluation of the presto combined qualitative real-time assay for Chlamydia trachomatis and Neisseria gonorrhoeae in Rwanda

Background: The Presto combined qualitative real-time assay for Chlamydia trachomatis and Neisseria gonorrhoeae (Presto CT/NG PCR assay) is appealing for developing countries, because it can be used with multiple DNA extraction methods and polymerase chain reaction (PCR) platforms.Objectives: The objective of the study was to implement and evaluate the Presto CT/NG PCR assay at the National Reference Laboratory (NRL) in Kigali, Rwanda, where no real-time PCR assays for the detection of C. trachomatis or N. gonorrhoeae were available.Methods: The Presto CT/NG PCR assay was first evaluated at the Institute of Tropical Medicine (ITM) in Antwerp, Belgium. Next, NRL laboratory technicians were trained to use the assay on their ABI PRISM 7500 real-time PCR instrument and their competencies were assessed prior to trial initiation. During the trial, endocervical swabs were tested at the NRL, with bi-monthly external quality control testing monitored by the ITM. The final NRL results were evaluated against extended gold standard testing at the ITM, consisting of the Abbott m2000 RealTime System with confirmation of positive results by an in-house real-time PCR assay for C. trachomatis or N. gonorrhoeae.Results: Of the 192 samples analysed using the Presto assay at the NRL, 16 samples tested positive for C. trachomatis and 17 tested positive for N. gonorrhoeae; four of these were infected with both. The sensitivity and specificity of the Presto assay were 93.3% (95% confidence interval [CI]: 68.1% ­ 99.8%) and 99.4% (95% CI: 96.8% ­ 100%) for C. trachomatis and 100% (95% CI: 76.8% ­ 100%) and 98.8% (95% CI: 95.8% ­ 99.9%) for N. gonorrhoeae.Conclusion: C. trachomatis and N. gonorrhoeae testing with the Presto assay was feasible in Kigali, Rwanda, and good performance was achieved

Evaluation of Reverse Hybridization Diagnostic Strategy for Simultaneous Detection of Chlamydia Trachomatis and Neisseria Gonorrhoeae

We performed a non radioactive reverse hybridization assay for simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae ; two pathogens influencing humain immunodeficiency virus (HIV) sexual transmission ; using 200 endo-cervical samples from pregnant women (Institute Pasteur n Cote d'Ivoire ).Diagnosis DNA sequences were amplified by multiplex polymerase chain reaction (PCR)using biotinylated primers. Amplified products were hybridized to different specific probes fixed as a line on membrane strips and visualized using a straptavidine /alkaline phosphatase conjugated system. The results were compared to Elisa (Enzyme linked Immunosorben Assay ) techniques and culture .The overall prevalence was 5and 15for N. gonerrhoeae and C. tranchomatis respectively and specificity of the reserve hybridization assay was high(100) .Additionally; PCR and radioactive hybridization reported here showed an increase in the sensitivity for detection of trichomonas vaginalis over conventional methods .The multiplex assays were highly effective for simultaneous detection of C. tranchomatis and N.gonorrhoeae in endocervical swab samples from women with without clinical signs of genitourinary infection .This technique allowing simultaneous diagnosis of pathogens could be great importance for epidemiological studies and management in public health interventions

Antimicrobial Susceptibility Patterns of Gonococcal Isolates in Pretoria; South Africa; over a 20-Year Period (1984-2004)

This paper reviews the susceptibility profiles of Neisseria gonorrhoeae over a 20-year period in the Pretoria region. Endourethral specimens were collected from adult men with symptoms of urethritis attending primary health care clinics and private medical practitioners. These swabs were plated on enriched media for isolation of N. gonorrhoeae. Antimicrobial susceptibility of the organisms was performed using the disc diffusion and agar dilution methods. Plasmid analyses were performed on beta-lactamase-producing isolates. Penicillase-producing N. gonorrhoeae strains increased from 4to 16; whilst chromosomally mediated penicillin-resistant strains increased dramatically from 0to 16from 1984 to 2004. There was an equal distribution of the 3.2 MDa African and 4.4 MDa Asian plasmids. High-level tetracycline-resistant strains (36) were detected for the first time in 2004. Ciprofloxacin resistance emerged at 7in the same year. Gonococcal isolates remained susceptible to cefoxitin; ceftriaxone; cefpodoxime; and spectinomycin. However; the minimum inhibitory concentration values for spectinomycin were very close to the breakpoint. We have shown a continuing increase in resistance to penicillin (plasmid and chromosomal); the emergence of high-level tetracycline resistance and an emergence of resistance to ciprofloxacin. Susceptibility testing is essential for successful therapeutic outcomes and needs to be performed in an ongoing basis

Antimicrobial Susceptibility Patterns of Gonococcal Isolates in Pretoria; South Africa; Over a 20-Year Period (1984-2004)

Abstract:This paper reviews the susceptibility profiles of Neisseria gonorrhoeae over a 20-year period in the Pretoria region. Endourethral specimens were collected from adult men with symptoms of urethritis attending primary health care clinics and private medical practitioners. These swabs were plated on enriched media for isolation of N. gonorrhoeae. Antimicrobial susceptibility of the organisms was performed using the disc diffusion and agar dilution methods. Plasmid analyses were performed on beta-lactamase-producing isolates. Penicillase-producing N. gonorrhoeae strains increased from 4 to 16; whilst chromosomally mediated penicillin-resistant strains increased dramatically from 0 to 16 from 1984 to 2004. There was an equal distribution of the 3.2 MDa African and 4.4 MDa Asian plasmids. High-level tetracycline-resistant strains (36) were detected for the first time in 2004. Ciprofloxacin resistance emerged at 7 in the same year. Gonococcal isolates remained susceptible to cefoxitin; ceftriaxone; cefpodoxime; and spectinomycin. However; the minimum inhibitory concentration values for spectinomycin were very close to the breakpoint. We have shown a continuing increase in resistance to penicillin (plasmid and chromosomal); the emergence of high-level tetracycline resistance and an emergence of resistance to ciprofloxacin. Susceptibility testing is essential for successful therapeutic outcomes and needs to be performed in an ongoing basis

Superior Detection of Pathogens in Synovial Fluid by the Bactec 9240 Peds Plus/F System Compared to the Conventional Agar-Based Culture Method

Abstract:To improve culture yield in cases of possible septic arthritis; we compared culture of joint fluid aspirates on conventional agar-based media to culture in Bactec 9240 Peds/Plus F blood culture bottles with and without the addition of fastidious organism supplement (FOS). Over a period of 21 months; we analysed 123 synovial fluid samples and isolated 20 pathogens. The Bactec methods proved superior by yielding more pathogens than the conventional culture method (p=0.074). However; this method also yielded more contaminants within the first three days of incubation (p=0.027). All contaminants detected after three days of incubation were the result of overgrowth on conventional method agar plates. The Bactec methods provided clinicians with a positive pathogen result one day earlier than the conventional counterpart (p=0.001). Four isolates of Neisseria gonorrhoeae were only cultured with the Bactec method. No significant benefit was demonstrated by supplementing blood culture bottles with FOS. We recommend that whenever infection by fastidious organisms is suspected; synovial fluid aspirates should be cultured using automated blood culture systems to increase the culture yield and to decrease the time to detection

Evaluation du premier jet d'urine dans la detection de Neisseria gonorrhoeae chez des patients paucisymptomatiques a Abdidjan (Cote d'Ivoire)

Il existe differentes methodes de diagnostic sur differents types de prelevement pour mettre en evidence Neisseria gonorrhoeae. Le prelevement de premier jet d'urine presente l'avantage d'etre non invasif; facilement reproductible et est deja utilise dans la detection de N. gonorrhoeae par des techniques moleculaires. L'objectif de cette etude etait d'evaluer le premier jet d'urine dans la detection de N. gonorrhoeae par les methodes classiques de bacteriologie chez des patients paucisymptomatiques. Elle a porte sur les prelevements de premier jet d'urine et de secretions uretrales de 87 patients de sexemasculin recus pour le diagnostic etiologique d'un syndrome d'infections sexuellement transmissibles. Un examen direct du frottis colore par la technique de Gram et des cultures sur le milieu de Thayer et Martin modifie et sur gelose au sang cuit enrichie en polyvitamines ont ete realises sur chacun des echantillons. La frequence des uretrites etait de 58;0. La gonococcie representait 7;5des cas. Le premier jet d'urines avait une sensibilite de 85;7et une specificite de 97;5dans la mise en evidence des diplocoques Gram negatif a l'examen direct. Par contre; il etait moins sensible que le prelevement de secretions uretrales dans la mise en evidence des uretrites microscopiques (sensibilite de 44;4et specificite de 100) et de la flore uretrale d'accompagnement (sensibilite de 59et specificite de 96;9). Les bonnes performances du premier jet d'urines dans la mise en evidence microscopique des diplocoques Gram negatif pourraient justifier son utilisation dans la detection deNeisseria gonorrhoeae dans un laboratoire de niveau 1. Il pourrait egalement etre utilise dans les etudes epidemiologiques et dans les enquetes de depistage a grande echelle

Prévalence et sensibilité aux antibiotiques de Neisseria gonorrhoeae chez les professionnelles du sexe et les patients IST au Niger

Le but de l'etude etait d'estimer la prevalence des infections a Neisseria gonorrhoeae et d'evaluer la sensibilite aux antibiotiques des souches; en vue d'une revision eventuelle du traitement par approche syndromique des ecoulements genitaux au Niger. De juillet a decembre 2007; un depistage systematique a ete effectue chez des professionnelles du sexe (PS) dans 5 localites; et des patients consultant pour IST a Niamey. Parallelement a l'examen microscopique apres coloration de Gram; les prelevements endocervicaux ou uretraux ont ete mis en culture sur gelose chocolat Polyvitex (bioMerieux); avec/sans inhibiteurs VCN; en atmosphere humide; sous 5-10CO2; pendant 1 a 2 jours. Les colonies suspectes sont identifiees sur galerie API NH (bioMerieux) et les antibiogrammes realises. Le gonocoque n'a ete isole que chez 0;68(5/739) des PS. Aucune souche n'a ete obtenue chez les 20 patientes IST; contre 2 chez les 9 cas d'uretrite masculine. Chacune des souches est resistante a au moins un des antibiotiques recommande par l'OMS dans l'approche syndromique. Cinq produisent une penicillinase et 2 une beta-lactamase a spectre elargi. Avec cette relative rarete des isolats et l'existence de resistance a au moins un des antibiotiques preconises; seule une surveillance microbiologique de routine permettra une collecte suffisante de souches pour avoir des donnees fiables necessaires a une revision de protocole

Antimicrobial suseptibility of neisserria gonorrhoeae isolated from patients attending various clinics in Pambeguwa, Kaduna State

Abstract: Antimicrobial resistant strains of gonococcal isolates have spread in many developing countries. In most African countries, the susceptibility of Neisseria gonorrhoeae is rarely tested. The antimicrobial susceptibility of one hundred and eight (108) Neisseria gonorrhoeae was therefore determined. Of the 108 isolates tested, 80 (74.07%) were resistant to penicillin, 74 (68.5%) were resistant to ampicillin. The resistance profiles for the remaining antibiotics tested were as follows: Erythromycin (58.3%), Tetracycline (62%) and gentamicin (25.9%). All the isolates were susceptible to ceftriaxone, cefuroxime and ofloxacin. Of the 80 penicillin resistant strains, 65 (81.3%) were penicillinase producing Neisseria gonorrhoeae (PPNG). The Minimum Inhibitory Concentration (MIC) of Penicillin against 9 gonococcal isolates, Ampicillin against 5 Neisseria gonorrhoeae isolates and Tetracycline against 2 gonococcal isolates was 32.0µg/ml. A total of 108 isolates were susceptible to 0.125µg/ml of ceftriaxone, cefuroxime and ofloxacin respectively. The high prevalence of PPNG and multiple drug resistance strains of Neisseria gonorrhoeae is associated with self-medication and sexual interaction between commercial sex workers (CSWs) resident in Pambeguwa and the long-distance truck drivers. Antimicrobial susceptibility test should be carried out on every gonococcal isolate obtained to ensure effective therapy. The present study was undertaken to assess the antimicrobial susceptibility of gonococcal isolates recovered from patients in Pambeguwa,Kaduna state, Nigeria

Prevalence of Sexually Transmitted Infections in Women Attending Antenatal Care in Tete Province; Mozambique

Objective. To determine the prevalence of Chlamydia trachomatis (CT); Neisseria gonorrhoeae (NG) and syphilis in pregnant women. Methods. A cross-sectional study was conducted among women attending antenatal care clinics (ANCs). Blood samples were tested for syphilis using the rapid plasma reagin (RPR) and treponemal haemaggluti- nation (TPHA) tests; CT and NG were diagnosed using a manual polymerase chain reaction assay on first-void urine samples. A socio-demographic questionnaire was completed. Results were compared with previous published data on sexually transmitted infection (STI) prevalence in Mozambique. Results. Blood and urine samples were collected from 1 119 and 835 women; respectively. The prevalence of CT was 4.1; and that of NG 2.5. The RPR test was positive in 5.2of the women; and 7.1had a positive TPHA test. Active syphiliswas found in 4.7. In univariate analysis; CT was associated with having had any level of education (p0.05); reactive RPR and TPHA were associated with illiteracy (p0.05); and TPHA was associated with age 25. Multivariate analysis did not show any significant association. In comparisonwith published data from 1993; a decline was observed for CT (p0.05); NG and syphilis (p0.001). Conclusions. Compared with available data; a decline of STI prevalence was observed in our setting. This might be the result of community-based education programmes focusing on changes to sexual behaviour; as well as the widespread use of the syndromic approach to managing STIs and the expansion of syphilis screening in primary health care settings. However; STI rates are still high; and the problem needs more concrete and sustained efforts for its control

A Study of Surface Proteins; Other Adhesins and Iron Acquiring Mechanisms in the Pathogenesis of Neisseria gonorrhoea strains Isolated from STD Patients in Ethiopia

The prevalence of Neisseria gonorrhoeae surface antigens as virulence factors in patients with sexually transmitted disease (STD) was determined. In addition; the potential of viral surface components as vaccine candidates against gonococcal infections was assessed. 335 urogenital specimens from male and female STD patients in Addis Ababa; Nazareth and Awassa; Ethiopia; were examined for N. gonorrhoeae. The virulence type; antimicrobial sensitivity pattern and surface antigenic components of isolated N. gonorrhoeae strains were determined using haemagglutination and other detection assay methods. Out of 335 specimens; N. gonorrhoeae strains were isolated in 59 of males and 43 of females. These strains showed 19 different antibiogram patterns. Only 12.3 of strains were sensitive to all drugs and over 70 were resistant to more than 2 combined drugs. More than 90 of these multidrug resistant strains were penicillinase [beta-lactamase] producers. 200 N. gonorrhoeaeisolates were tested for haemagglutination and iron-binding protein production (IBP). Results showed that 35 of tested isolates were mannose resistant haemagglutinating (MRHA) strains and 20 were mannose sensitive haemagglutinating (MSHA) strains when tested using the human AB blood type. When guinea pig blood cells were used; 15 were MRHA and 25 were MSHA strains. MRHA strains were positive for IBP; whereas only 10 of MSHA strains were positive for this property. Conclusions and recommendations are given

Antimicrobial susceptibility of Neisseria gonorrhoea in Kumasi

Between January - December 1992; three hundred and fifty six (356) isolates of Neisseria gonorrhoeae from clinical material were subjected to antimicrobial susceptibility testing in the Clinical Microbiology Laboratory of the Komfo Anokye Teaching Hospital; Kumasi. Ceftriaxone; Cefuroxime; and Norfloxacin showed high activity against the gonococcus (95-100 per cent sensitivity). Seventy-eight per cent (78 per cent) of the isolates were sensitive to Spectinomycin. a low activity was exhibited by the commonly used antimicrobials; i.e. Penicillin 8.5 per cent; Tetracycline 2.5 per cent; Cotrimozale 3 per cent; Erythromycin 8.5 per cent; and Chloramphenicol 10 per cent

Subacute and acute epidemic conjunctivitis due to neisseria gonorrhoeae SSP

An outbreak of acute conjunctivitis with profuse exudation was observed between November 1987 and June 1988 in Konso; North Omo. There were 9;075 registered cases and seven months later; 20 cases with mild; subacute conjunctivitis were observed. Neisseria gonorrhoeae and Neisseria gonorrhoeae subsp. kochii were the causative agents; while Haemophilus spp. behaved as opportunistic infection. The condition is resistant to topical antibiotics; but the outbreak could be controlled by mass treatment with single dose oral ampicillin and probenecid. Severe sequelae were observed in 36 patients treated with this regimen. This is the first isolation of N. gonorrhoeae subsp. kochii outside Egypt; and the first recorded outbreak of Neisseria gonorrhoeae conjunctivitis affecting thousands of patients in more than 40 years

Prevalence et antosensibilite des gonoccoques isoles a Niamey : analyse de 6.608 prelevements genitaux

Un bilan de la recherche de neisseria gonorrhoeae dans 6.608 prelevements genitaux effectues de 1985 a 1992; a Niamey est presente. Ce germe a ete isole 624 fois; soit une prevalence globale de 9;44 pour cent. Quant aux antibiogrammes; pour les 17 antibiotiques retenus; l'on constate de grandes variations dans la frequence de leur utilisation. Cela n'a pas permis de tirer des conclusions generalisables sur les taux d'efficacite de chacun de ces produits sur le gonocoque. Celui-ci semble cependant plus frequemment sensible au chloramphenicol; aux cyclines et a l'oleandomycine