Comparison of two methods for detection of Chlamydia trachomatis and Ureaplasma urealyticum in male reproductive tract / 北京大学学报(医学版)

OBJECTIVE@#To investigate the value of clinical application of simultaneous amplification and testing of RNA (SAT-RNA) for detecting Chlamydia trachomatis (CT) and Ureaplasma urealyticum (UU) by comparing with the polymerase chain reaction testing of DNA (PCR-DNA) method.@*METHODS@#Specimens from both urethra swab and the first avoid urine which should be at least one hour after the previous urination were collected from 163 men who were scheduled for in vitro fertilization and embryo transfer (IVF-ET) treatment due to female factors at Center for Reproductive Medicine, Shengjing Hospital of China Medical University during the period of April 2016 to April 2017. Among the 163 men, 109 simultaneously provided semen that was collected after 3-7 days of sexual abstinence for the testing. Urine and semen specimens were detected for CT and UU with SAT-RNA, while urethra swab specimens were detected for CT and UU with standard PCR-DNA. Detection results of the SAT-RNA were compared with those of the PCR-DNA method.@*RESULTS@#The positive rate of UU in the urethra swab detected with PCR-DNA and that of UU in the urine with SAT-RNA were 47.24% and 47.85%, respectively, and the coincidence rate was 93.25%. In addition, the positive and negative coincidence rates were 93.51% and 93.02%, respectively, and the concordance between the two methods was very good (Kappa=0.865). On the other hand, the positive rate of CT in the swab specimen tested with PCR-DNA was 3.07% and that of CT in urine with SAT-RNA was 4.29%, and the coincidence rate was 97.55%. Moreover, the positive and negative coincidence rates were 80.00% and 98.10%, respectively, and the concordance between the two methods was good (Kappa=0.654). Regarding SAT-RNA detection of UU in the urine and semen specimen of the 109 patients, the positive rates of UU in the urine and semen specimens were 50.46% and 44.95%, respectively; and the coincidence rate between the two specimens was 88.99%. In addition, the positive coincidence rate and the negative coincidence rate was 93.88% and 85.00%, respectively, and the concordance between the two specimens was good (Kappa=0.780). Similarly, SAT-RNA detection of CT in the urine and semen specimens showed the positive rate was 5.50% and 3.67%, respectively; and the two specimens showed 98.17% coincidence rate. The positive and negative coincidence rates were 100.00% and 98.10%, respectively, and the concordance was also good (Kappa=0.791).@*CONCLUSION@#SAT-RNA detection of CT and UU in the urine specimen showed good concordance with the PCR-DNA detection of CT and UU in the urethra swab specimen. In addition, the concordance was also good between the urine and semen specimens detected with SAT-RNA. These results indicate that, as a less invasive and equally accurate procedure, SAT-RNA may be more suitable for clinical application.

Temporal evolution of antimicrobial resistance among Neisseria gonorrhoeae clinical isolates in the most populated South American Metropolitan Region

A total of 124 Neisseria gonorrhoeae isolates recovered during a 12-year period (2003-2015) from outpatients assisted at Centro de Referência e Treinamento DST/AIDS-CRT of São Paulo city, Brazil, were analysed. The following resistance rates were observed: penicillin-59.6%, ciprofloxacin-15.3%, and azithromycin-6.7%. Although reduced susceptibility to these drugs was observed since 2003, no ceftriaxone-resistant isolates were detected. Ciprofloxacin- and azithromycin non-susceptible isolates were grouped in 11 clusters. Mutations were detected in GyrA and ParC of isolates 124 and 260, and a C2611T substitution on 23S rRNA alleles was also observed in isolate 260. Both isolates belonged to ST1901/ST6210 (MSLT/NG-MAST schemes).

Antimicrobial resistance in Neisseria gonorrhoeae: history, molecular mechanisms and epidemiological aspects of an emerging global threat

ABSTRACT Neisseria gonorrhoeae is the agent of gonorrhea, a sexually transmitted infection with an estimate from The World Health Organization of 78 million new cases in people aged 15-49 worldwide during 2012. If left untreated, complications may include pelvic inflammatory disease and infertility. Antimicrobial treatment is usually effective; however, resistance has emerged successively through various molecular mechanisms for all the regularly used therapeutic agents throughout decades. Detection of antimicrobial susceptibility is currently the most critical aspect for N. gonorrhoeae surveillance, however poorly structured health systems pose difficulties. In this review, we compiled data from worldwide reports regarding epidemiology and antimicrobial resistance in N. gonorrhoeae, and highlight the relevance of the implementation of surveillance networks to establish policies for gonorrhea treatment.

Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum, and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction

The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period.

Nucleic acid amplification tests (NAATs) for gonorrhoea diagnosis in women: Experience of a tertiary care hospital in north India.

Background & objectives: Gonorrhoea is among the most frequent of the estimated bacterial sexually transmitted infections (STIs) and has significant health implications in women. The use of nucleic acid amplification tests (NAATs) has been shown to provide enhanced diagnosis of gonorrhoea in female patients. However, it is recommended that an on-going assessment of the test assays should be performed to check for any probable sequence variation occurring in the targeted region. In this study, an in-house PCR targeting opa-gene of Neisseria gonorrhoeae was used in conjunction with 16S ribosomal PCR to determine the presence of gonorrhoea in female patients attending the tertiary care hospitals. Methods: Endocervical samples collected from 250 female patients with complaints of vaginal or cervical discharge or pain in lower abdomen were tested using opa and 16S ribosomal assay. The samples were also processed by conventional methods. Results: Of the 250 female patients included in the study, only one was positive by conventional methods (microscopy and culture) whereas 17 patients were found to be positive based on PCR results. Interpretation & conclusions: The clinical sensitivity of conventional methods for the detection of N. gonorrhoeae in female patients was low. The gonococcal detection rates increased when molecular method was used giving 16 additional positives. Studies should be done to find out other gene targets that may be used in the screening assays to detect the presence of gonorrhoea.

Experiencia con un método de autotoma de muestra vaginal para la detección de infección por Chlamydia trachomatis y Neisseria gonorrhoeae en mujeres jóvenes / Screening of Neisseria gonorrhoeae and Chlamydia trachomatis using techniques of self collected vaginal sample in young women

Chlamydia trachomatis and Neisseria gonorrhoeae are responsible for 3-10% of sexually transmitted diseases in adolescents. 75% are asymptomatic. International standards recommend annual screening for C. trachomatis in sexually active women under 26 years. Self-collected vaginal swab is one of the less invasive screening methods, it is well accepted by patients and rarely used in our country. Aim: To determine the frequency of C. trachomatis and N. gonorrhoeae by a self-sampling method of vaginal swab and its acceptability in a group of adolescents and young adults. Patients and Methods: Women 18 to 25 years old. Vaginal samples were processed by nucleic acid amplification tests, Gen Probe APTIMA Combo2. Data were collected on sexual behavior and perception of self-sampling by survey. Results: We studied 344 patients with an average age of 21.7 years. Detection of C. trachomatis was positive in 7.9% women and it was not found in any of the patients studied for N. gonorrhoeae. 98% considered self-sampling instructions easy to understand, 87.5% felt comfortable taking the sample. Conclusions: Prevalence of C. trachomatis in the study population was similar to that described in other national and international studies. N. gonorrhoeae was not found in this series, which is consistent with literature reports. The self-sampling technique of vaginal sample was well accepted by the patients. However, they were anxious about the quality of the sample. According to our results, it is important to emphasize the importance of annual detection of these pathogens and that self-sampling technique is a valid alternative.

Chlamydia trachomatis y Neisseria gonorrhoeae son causantes de 3 a 10% de las infecciones de transmisión sexual en adolescentes. Las normas internacionales recomiendan su detección anual en mujeres sexualmente activas menores de 26 años. La adherencia a este tamizaje en mujeres jóvenes está limitada por el temor al examen ginecológico y alto costo del examen. Objetivo: Determinar la frecuencia de detección de C. trachomatis y N. gonorrhoeae por un método de auto-toma de muestra vaginal y su aceptabilidad en un grupo de adolescentes y jóvenes adultas. Pacientes y Método: Se incluyeron mujeres de 18 a 25 años atendidas en Clínica Las Condes y el Servicio de Salud Estudiantil de la Universidad de Chile, que fueron instruidas para autotoma de muestra vaginal. Luego de dar su consentimiento, las muestras fueron estudiadas mediante reacción de polimerasa en cadena para la detección de C. trachomatis y N. gonorrhoeae. Se recopilaron datos sobre conductas sexuales y percepción de la autotoma mediante encuesta. Se determinó la relación entre estos factores y la aceptabilidad del método. Resultados: Se reclutaron 344 mujeres, con una edad promedio de 21,7 años. La detección de C. trachomatis fue de 7,9% y no se encontró muestra positiva para N. gonorrhoeae. El reporte de flujo vaginal por la paciente se asoció a 1,5 veces mayor riesgo de C. trachomatis. El 98% consideró las instrucciones de la autotoma fáciles de entender, 87,5% se sintió cómoda al tomar la muestra. Conclusiones: La prevalencia de C. trachomatis en la población estudiada fue similar a lo descrito en otras series nacionales e internacionales; no se encontró N. gonorrhoeae en esta serie, lo que coincide con lo reportado en el extranjero. La técnica de autotoma de muestra vaginal fue bien aceptada por las pacientes; sin embargo, manifestaron ansiedad acerca de la seguridad de una toma adecuada. De acuerdo a nuestros resultados, es importante insistir en la detección anual de estos patógenos siendo la técnica de autotoma una alternativa válida.

Antimicrobial susceptibility of Neisseria gonorrhoeae isolates from patients attending a public referral center for sexually transmitted diseases in Belo Horizonte, State of Minas Gerais, Brazil

Introduction The aim of this study was to determine the antimicrobial susceptibility of Neisseria gonorrhoeae isolates obtained from patients attending a public referral center for sexually transmitted diseases and specialized care services (STD/SCS) in Belo Horizonte, Brazil. Methods Between March 2011 and February 2012, 201 specimens of Neisseria gonorrhoeae were consecutively obtained from men with symptoms of urethritis and women with symptons of cervicitis or were obtained during their initial consultation. The strains were tested using the disk diffusion method, and the minimum inhibitory concentrations of azithromycin, cefixime, ceftriaxone, ciprofloxacin, chloramphenicol, penicillin, tetracycline and spectinomycin were determined using the E-test. Results The specimens were 100% sensitive to cefixime, ceftriaxone and spectinomycin and exhibited resistances of 4.5% (9/201), 21.4% (43/201), 11.9% (24/201), 22.4% (45/201) and 32.3% (65/201) to azithromycin, ciprofloxacin, chloramphenicol, penicillin and tetracycline, respectively. Intermediate sensitivities of 17.9% (36/201), 4% (8/201), 16.9% (34/201), 71.1% (143/201) and 22.9% (46/201) were observed for azithromycin, ciprofloxacin, chloramphenicol, penicillin and tetracycline, respectively. The specimens had plasmid-mediated resistance to penicillin PPNG 14.5% (29/201) and tetracycline TRNG 11.5% (23/201). Conclusions The high percentage of detected resistance to penicillin, tetracycline, chloramphenicol and ciprofloxacin indicates that these antibiotics are not appropriate for gonorrhea treatment at the Health Clinic and possibly in Belo Horizonte. The resistance and intermediate sensitivity of these isolates indicates that caution is recommended in the use of azithromycin and emphasizes the need to establish mechanisms for the surveillance of antimicrobial resistance for the effective control of gonorrhea. .

Molecular basis of high-level ciprofloxacin resistance in Neisseria gonorrhoeae strains from Shandong Province, China

In the study, the ciprofloxacin resistance rate was 100%. High-level ciprofloxacin resistance rate was 63.55%. Sixteen different mutation patterns involved in the formation of ciprofloxacin resistance were identified. The most prevalent were patterns P7 (25.2%), P8 (15.0%), P9 (11.2%), P1 (10.3%), and P5 (10.3%). All of the 107 NG isolates analyzed for mutations in the study have demonstrated a change of Ser-91 → Phe in the gyrA gene, and all except one have demonstrated a change in position 95 of the amino acid sequence. All of the 68 high-level QRNG isolates had double mutations in gyrA gene combined with a single or two mutations in parC gene. It is most important that a new mutation site of Ile-97 → Met in gyrA and a new mutation of Leu-106 → Ile in parC were found in the study, both leading to high-level ciprofloxacin resistance (MIC values, 8 µg/mL, 32 µg/mL, respectively). Therefore, we confim that gyrA mutations are necessary for the fluoroquinolone resistance phenotype and parC mutations are correlated intimately with high-level fluoroquinolone resistance. In China fluoroquinolone resistance in Neisseria gonorrhoeae strains is very serious and the new mutation sites in the fluoroquinolone resistance-determining regions emerge more and more quickly. Hence, in China fluoroquinolones, which are used to treat gonorrhoea presently, should be substituted by a new antibiotics.

An insight into the drug resistance profile & mechanism of drug resistance in Neisseria gonorrhoeae.

Among the aetiological agents of treatable sexually transmitted diseases (STDs), Neissseria gonorrhoeae is considered to be most important because of emerging antibiotic resistant strains that compromise the effectiveness of treatment of the disease - gonorrhoea. In most of the developing countries, treatment of gonorrhoea relies mainly on syndromic management rather than the aetiological based therapy. Gonococcal infections are usually treated with single-dose therapy with an agent found to cure > 95 per cent of cases. Unfortunately during the last few decades, N. gonorrhoeae has developed resistance not only to less expensive antimicrobials such as sulphonamides, penicillin and tetracyclines but also to fluoroquinolones. The resistance trend of N. gonorrhoeae towards these antimicrobials can be categorised into pre-quinolone, quinolone and post-quinolone era. Among the antimicrobials available so far, only the third-generation cephalosporins could be safely recommended as first-line therapy for gonorrhoea globally. However, resistance to oral third-generation cephalosporins has also started emerging in some countries. Therefore, it has become imperative to initiate sustained national and international efforts to reduce infection and misuse of antibiotics so as to prevent further emergence and spread of antimicrobial resistance. It is necessary not only to monitor drug resistance and optimise treatment regimens, but also to gain insight into how gonococcus develops drug resistance. Knowledge of mechanism of resistance would help us to devise methods to prevent the occurrence of drug resistance against existing and new drugs. Such studies could also help in finding out new drug targets in N. gonorrhoeae and also a possibility of identification of new drugs for treating gonorrhoea.

A Case of Bacteremia by Neisseria gonorrhoeae Coincident with Massive Hemorrhage of Esophageal Varices / 대한진단검사의학회지

A 42-yr-old man with hepatitis B virus associated liver cirrhosis was admitted to the emergency room because of multiple seizures, a history of chills and myalgia over the previous 2 weeks, and 3 days of melena. He was febrile with a temperature of 38.0degrees C. There were no symptoms and signs related to the genitourinary system, skin, or joints. Three sets of blood cultures were obtained and oxidase-positive, gram-negative diplococci were detected after 25.9-26.9 hr of incubation in all aerobic vials. The organism was positive for catalase and oxidase, and was identified as Neisseria gonorrhoeae, using a Vitek Neisseria-Haemophilus Identification card (bioMerieux Vitek, Inc., USA). Further, 16S rRNA sequencing of this isolate revealed a 99.9% homology with the published sequence of N. gonorrhoeae strain NCTC 83785 (GenBank Accession No. NR_026079.1). Acute bleeding by variceal rupture seems to be a likely route of introduction of N. gonorrhoeae from the mucosa into the blood. To the best of our knowledge, this is the first case of gonococcal bacteremia in Korea.

Fluoroquinolone resistance among Neisseria gonorrhoeae isolates from Shanghai, China: Detection of quinolone resistancedetermining region mutations.

Background & objectives: Fluoroquinolone has a broad spectrum of antimicrobial activity, and is widely used for gonorrhoea treatment. However, its effi cacy can be compromised by the drug-resistance property of Neisseria gonorrhoeae isolates. Most resistant cases of N. gonorrhoeae are associated with mutations in the quinolone-resistance-determining-region (QRDR) within genes of gyrA and parC. This study was undertaken to describe resistance profi le of N. gonorrhoeae to fl uoroquinolones in Shanghai, P.R. of China, and also associated resistance mutations in gyrA and parC. Methods: Eighty N. gonorrhoeae isolates were collected from Shanghai Skin Disease & Sexually Transmitted Disease Hospital or DongFang Hospital during April 2005 to April 2006 in Shanghai, P.R. of China. The minimum inhibitory concentrations (MIC) of fl uoroquinolones for these isolates were determined by an agar dilution method. Mutation patterns within gyrA and parC were determined by direct sequencing or by using established restriction fragment length polymorphisms (RFLP) methods. Results: Ninety fi ve per cent (76 of 80) of isolates were resistant, 3.75 per cent (3 of 80) intermediate resistant, and 1.25 per cent (1 of 80) were sensitive to fl uoroquinolone drug ciprofl oxacin. Sequencing and RFLP analysis of gyrA and parC revealed that all resistant isolates had dual mutations of S91F and D95A/G/N in gyrA. Some isolates had an extra mutation within parC either of D86N, S87N or E91A/G. Mutation patterns for gyrA and parC were signififififi cantly (P<0.05) associated with MICs level. Interpretation & conclusions: Mutations of S91F and D95A/G/N in gyrA combined with S87N in parC was the most prevalent mutation pattern of fl uoroquinolone resistant N. gonorrhoeae isolates. This mutation pattern was associated with a high level of quinolone resistance (MIC >16.0 μg/ml) which can serve as a maker for quinolone-resistance prediction in Shanghai, P.R. of China.

In vitro recombination and identification of mutated fragment corresponding to regulation region of mtrR gene of Neisseria gonorrhoeae / 华中科技大学学报（医学）（英德文版）

A site-directed mutant DNA fragment was synthesized and transfected into clinical Neisseria gonorrhoeae (NG) stains to construct the transformants that contained the corresponding mutagenesis of regulation region of mtrR gene. According to the technique of gene splicing by overlap extension (SOEing), a DNA segment with specific mutagenesis was constructed by two-step polymerase chain reaction (PCR). The mutation fragments EF could be used for the next experiment in which the mutation NG strains were induced. By comparing the recombinant EF fragments to the corresponding DNA fragments of clinical NG strains, 2 of these were not compatible completely. The results of sequencing revealed that there was a 9 bp deletion between the 45 to 54 inverted repeat sequence localized within the mtrR promoter. It can be confirmed that the fragments EF are the specifically designed mutant fragments.

Screening for Chlamydia and Gonorrhea by Strand Displacement Amplification in Homeless Adolescents Attending Youth Shelters in Korea

We conducted the screening of sexually transmitted infections to define the prevalence of genital Chlamydia trachomatis and Neisseria gonorrhoeae infections and status of sexual risk behavior among homeless adolescents (10-19 yr old) in Korea. Adolescents who ran away from home and are under the care of youth shelters in ten cities in Korea served as the study population. Participants filled out a self-administered questionnaire related to sexuality. First-void urine was analyzed for chlamydial and gonococcal infection by strand displacement amplification (BDProbTec(TM)ET, BD Diagnostic Systems, MD, U.S.A.). A total of 175 adolescents from 15 youth shelters took part in the study. Their median age was 16 yr, and 54.9% of them reported having sexual intercourse at least once. The prevalence of C. trachomatis and N. gonorrhoeae among homeless adolescents was 12.6% and 15.4%, respectively. Factors significantly associated with the infections were number of sexual partners during the past year and lifetime. This is the first community-based sexually transmitted infection (STI) screening among adolescent in Korea. Screening programs targeting sexually active adolescents are important for detection of STIs. They should be considered an alternative population-based surveillance system in order to control STIs nationally.

Advances in diagnostic tests for bacterial STDs

Las pruebas de laboratorio son necesarias a menudo para el diagnóstico de las infecciones transmitidas sexualmente, debido a la naturaleza asintomática o a la presencia de síntomas inespecíficos de esas infecciones. En este sentido, durante los años relativamente recientes se han registrado importantes avances tecnológicos, como por ejemplo los ensayos de amplificación de ácidos nucleicos que han permitido una mejora en la posibilidad de diagnosticar las infecciones causadas por Chlamydia trachomatis. El descubrimiento de que las pruebas de amplificación de ácidos nucleicos permiten diagnosticar a un mayor número de individuos infectados y de que son útiles para tamizar poblaciones con bajas prevalencias de infección, han conducido al desarrollo de estrategias diseñadas para reducir el costo de los ensayos de laboratorio sin que ello impacte significativamente en la sensibilidad de las pruebas diagnósticas. Por otra parte, el desarrollo de nuevas pruebas para el diagnóstico de la sífilis ha ganado momento a partir de la factibilidad de producir un antígeno de VDRL sintético, que deberá resultar en mejores pruebas de anticuerpos no-treponémicos para el tamiz de la sífilis. Ahora bien, aún cuando se ha completado el conocimiento de la secuencia genética del Treponema pallidum, este microrganismo todavía no es susceptible de cultivarse in vitro. Sin embargo, la revolución de la biología molecular ha facilitado la implantación de ensayos de la reacción en cadena de la polimerasa para detectar al Treponema pallidum en varios tipos de muestras clínicas, así mismo ahora es posible la producción de antígenos recombinantes de esa bacteria para utilizarse en pruebas serológicas de anticuerpos treponémicos específicos. En conclusión, es de esperarse que la investigación futura favorecerá la disponibilidad de pruebas de laboratorio sensibles y de bajo costo para el diagnóstico de las infecciones transmitidas sexualmente.

Emerging antimicrobial resistance, plasmid profile and pulsed-field gel electrophoresis pattern of the endonuclease-digested genomic DNA of Neisseria gonorrhoeae

Resistant gonococci are very prevalent in many countries, particularly in Asia. This study was conducted to determine the trend of resistance, the effect of decreasing the ciprofloxacin susceptibilities of gonococci on the prevalence of penicillinase-producing N. gonorrhoeae (PPNG), and to compare the epidemiology of strains with the previous studies. A total of 602 strains of gonococci were isolated from prostitutes in 1997-1999. Antimicrobial susceptibility was tested by NCCLS disk diffusion and agar dilution methods. For epidemiologic analysis, vplasmid analysis and pulsed-field gel electrophoresis (PFGE) were performed. The proportion of PPNG remained high (79%), and the strains with decreased susceptibility to ciprofloxacin increased significantly from 67% in 1997 to 84% in 1999. Compared to our previous study, the PFGE patterns were similar, while the proportion of strain with the 3.2-MDa plasmid markedly decreased. In conclusion, a rapid increase in ciprofloxacin-nonsusceptible strains may suggest difficulties in the treatment of gonococcal infections in the near future with the drug. The recent decrease of PPNG with the 3.2-MDa plasmid may suggest that there is an epidemiological change in gonococcal infections, and the prevalence of related PFGE patterns suggests the dissemination of a few clones among the high risk populations.

Molecular characterization of plasmid content of penicillinase-and non-penicillinase-producing neisseria gonorrhoeae isolates and its epidemiologic correlate

A total number of 40 gonococcal strains isolated from patients were classified according to the production of beta-Iactamase [penicillinase] enzyme into 32 penicillinase-producing [PPNG] and 8 non- penicillinase-producing [non-PPNG] strains. Plasmid DNA was extracted from all 32 PPNG strains and 7 non-PPNG strains and was subjected to agarose gel electrophoresis. Twenty three PPNG strains showed an identical. plasmid profile composed of 3 different plasmid species having masses of 2.6-, 24.5, and 24.5 - Md, 5 PPNG strains carried only the 2.6- and 4.5- Md plasmid, whereas the remaining 4 PPNG strains harboured a 3.2- Md plasmid in addition to the 2.6- Md plasmid. Out of the 8 non-PPNG strains 7 carried the 2.6- Md plasmid, but only 1 carried the 24.5- Md plasmid in addition and I was plasmid- free. Epidemiologically, it was found that 20 [83%] of the 24 PPNG strains isolated from patients who contracted the gonococcal infection in the Far East carried the 3 different plasmids 2.6-, 4.5, and 24.5- Md, thus all contained the 4.5- Md [Asia] plasmid. The 4 PPNG strains isolated from men who contracted infection in Moroco contained the 2.6-Md plus the 3.2-Md [Africa] plasmid. On the other hand, the 4 PPNG strains isolated from men returning from Turkey had 2 different plasmid profiles, 3 strains carried the 3 different plasmid 2.6-, 4.5, and 24.5-Md and 1 strain carried the 2 different plasmids 2.6, and 4.5- MD

Actividad in vitro de agentes antimicrobianos selectos frente a cepas de Neisseria gonorrhoese, streptococcus pneumoniae y streptococcus beta - hemolítico / In vitro activity of selected antimicrobial agents ogeinst of neisseria gonorrhoeae, streftococcus pneumoniae and beta hemolytic streptococcus

Las acividades in vitro de diez agentes antimicrobianos incluyendo a la Penicilina, Ampicilina, Cefalotina, Imipenem, Tetraciclina, Eritromicina, Trimetoprim-Sulfametoxazole, Gentamicina, y un nuevo agente B-lactámico (Imipenem) y dos nuevas quinolonas (Norfloxacina y Ciprofloxacina), fueron comparadas contra 46 cepas de Neisseria gonorrhoeae, 19 cepas de Streptococcus pneumoniae y 16 cepas de Streptococcus B-hemolíticos (especie inepecífica, agrupadas por presentar un número muy pequeño por separado). Todas las cepas fueron aisladas de muestras clínicas recibidas en el laboratorio de Microbiología Clínica del Instituto de Medicina Tropical Alexander Von Humboldt de la Universidad Peruana Cayetano Heredia (Lima-Perú). La susceptibilidad antibacteriana fue estudiada por el método de difusión con disco (antibiograma) y por el método de agar dilución, a través de la determinación de la Concentración Inhibitoria Mínima (CIM). la Penicilina continúa exhibiendo una buena actividad in vitro contra la Neisseria Gonorrhoeae, y puede considerarse a la Cefalotina, al Imipenem y a las 2 fluoroquinolonas (Norfloxacina y Ciprofloxacina) como alternativas de tratamiento. Es necesario resaltar que la no susceptibilidad a la Penicilina se relaciona con la actividad B-lactamasa (mas), en forma significativa (p igual 0.0178). La mayoría de los agentes antimicrobianos presentaron una buena actividad in vitro frente a las cepas de Streptococcus Pneumoniae, exceptuando al Cotrimoxazole, a la Gentamicina y a la Tetraciclina. Se encuentra la aparición de cepas moderadamente susceptibles contra la Penicilina (con un CIM igual a 0.25 ug/ml). Las cepas de Streptococcus B-hemolíticos demostraron la menor susceptibilidad in vitro, siendo sólo susceptibles a los agentes B-lactámicos. En especial, las nuevas quinolonas no han presentado una actividad adecuada frente a estos microorganismos. Se debe promocionar un control periódico de la susceptibilidad antimicrobiana in vitro para detectar rápida y oportunamente la aparición de cepas no susceptibles, y para brindar un nuevo instrumento al médico clínico que lo ayude a tomar las decisiones para brindar el tratamiento más adecuado

Diagnosis of gonorrhoea using a genetic transformation test at a venereal disease clinic in Thailand.

A genetic transformation test (GTT), a technique in which gonococcal DNA is detected in clinical specimens, was used to search for Neisseria gonorrhoeae infections in 37 men and 159 women at the Venereal Disease clinic in Cholburi, Thailand. Swabs were collected in duplicate from cervical specimens from 159 women and from urethral specimens from 37 men. One of each specimen was cultured on Thayer-Martin media while the other was mailed to the United States at room temperature for the GTT which involved a delay of 10 to 14 days. With the urethral specimens N. gonorrhoeae was identified in 84% (31/37) of specimens and there was 100% concordance between the results of the GTT and culturing specimens directly on Thayer-Martin media. With cervical specimens N. gonorrhoeae was isolated from 26% (41/159) by the standard culture technique and 19% (13/159) by the GTT. Seventy-six percent of the culture positive specimens were positive with the GTT and two specimens from which N. gonorrhoeae were not isolated were positive in the GTT. The GTT technique enables physicians to send swab collected from patient with suspected gonorrhoea without any special transport media to a central laboratory for laboratory diagnosis of gonorrhoeal infections. This technique which uses reagents which are available in most bacteriology laboratories, should facilitate surveillance of gonorrhoea especially when specimens are collected in clinics where bacteriology laboratory facilities are not available.