Comparison of culture and real-time polymerase chain reaction methods for detection of Mycoplasma hominis in amniotic fluids samples.

BACKGROUND: Mycoplasma hominis is often present in the amniotic cavity with microbial invasion associated with spontaneous preterm labor. Conventional culture method is the gold standard for detection of Mycoplasmas, but real-time polymerase chain reaction (real-time PCR) has revolutionized the diagnosis of M. hominis. OBJECTIVE: The purpose of this study is the comparison of the culture methodology with real-time PCR for the detection of M. hominis in amniotic fluid samples. METHODS: Amniotic fluid samples were collected from 65 pregnant women (age range: 25-45 years) previously followed at an infertility clinic. They were collected by transabdominal genetic amniocentesis during 16-21 weeks of gestation. Amniotic fluids were inoculated in SP4 broth for 48-72 h, and after becoming alkaline, culture suspension was spread on A7 agar plate for 1 week till the typical colonies seen in "fried-egg" morphology under stereomicroscope. DNA was extracted using a QIAGEN Mini DNA kit. The real-time-PCR was performed using Rotor-Gene Q Real-time PCR instrument. A melting-curve analysis was also performed. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were measured by real-time PCR by taking culture as gold standard. RESULTS: Sixty-five women in 16-21 weeks of gestation, with a mean age of 33 ± 5.06 years, were enrolled into this study. M. hominis detected by culture and real-time PCR assay was 72% (47/65) and 69% (45/65), respectively. 66% (43/65) specimens were positive by both methods. Real-time PCR sensitivity was 91.5%, specificity 88.9%, PPV 95.6%, and NPV 80%. CONCLUSION: Rapid detection of Mycoplasmas causing maternal complications such as neonatal infections and preterm labor in pregnancy by real-time PCR may be important and necessary. The high sensitivity and shorter time requirement of real-time PCR support its further development for diagnosis of Mycoplasma infections.

Prevalence of Human Herpesvirus-8 and BK Polyoma Virus Infections in End-stage Renal Disease and the Influence of Renal Transplantation.

Viral infections lead to significant morbidity and mortality in kidney transplant recipients. We evaluated 49 kidney transplant recipients for human herpesvirus 8 (HHV-8) and BK polyomavirus infections in conjunction with data obtained from 43 donors. The seroprevalence of HHV-8 was 6.9% in donors and 12.2% in recipients. HHV-8 DNA was detected below the limit of quantification (<5000 copies/mL) in a recipient with HHV-8 seropositivity at the pretransplant period and was undetectable at month 3 after transplantation. Transient viruria with BK polyomavirus was recorded in 10.2% of recipients without viremia. Multiple factors contribute to viral reactivation, particularly immunosuppressive treatment. Reduction in maintenance immunosuppression seems beneficial in terms of viral reactivation. At our center, routine use of valganciclovir for antiviral prophylaxis may be effective for the prevention of HHV-8 reactivation.

Steroid-refractory inflammatory bowel disease is a risk factor for CMV infection.

OBJECTIVE: Patients with inflammatory bowel disease (IBD) show increased the prevalence of cytomegalovirus (CMV) infection due to the severity of the disease and the immunosuppressive treatments they receive. The aim of this study was to determine the prevalence of CMV infection in IBD patients and identify the risk factors for CMV infection with different demographic characteristics in IBD patients. PATIENTS AND METHODS: We enrolled 85 patients diagnosed with IBD (43 with ulcerative colitis (UC) and 42 with Crohn's disease (CD)) in this prospective study. The clinical disease activities of UC and CD were assessed using Truelove-Witts and Crohn's disease activity index (CDAI). CMV infection was assessed by detection of DNA using real-time polymerase chain reaction (PCR) in blood samples and quantitative PCR in colonic biopsy specimens and by detection of inclusion bodies using hematoxylin-eosin staining. RESULTS: Thirteen patients with IBD exhibited concomitant CMV infection. CMV infection was not detected in any of the patients in remission. Viral loads measured in the colonic mucosa of infected patients ranged from 800-7000 genome copies/mL total extracted DNA. The mean serum CMV DNA level was 1694 ± 910 copies/mL (range: 800-3800). The rate of steroid resistance in CMV-positive cases was significantly higher than that in CMV-negative cases (p = 0.001). CD with acute exacerbation was a risk factor for CMV disease (p = 0.04). All of the CMV-positive patients received immunosuppressive treatments. CONCLUSIONS: CMV infection should be suspected in steroid-resistant UC and CD. Antiviral treatment improved the clinical outcome in steroid-resistant IBD cases with serum CMV DNA levels above 1000 copies/mL.

Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae in Turkey among men With urethritis.

BACKGROUND: Chlamydia trachomatis and Neisseria gonorrhoeae are known to cause urethritis. However, only a small number of studies in Eastern European countries have investigated the causes of urethritis. GOALS: To determine the prevalence of C trachomatis and N gonorrhoeae among men with symptomatic urethritis in Istanbul, Turkey, and to determine whether contact with a commercial sex worker increased the likelihood of chlamydial infections. STUDY DESIGN: Men with a diagnosis of urethritis at the Istanbul Faculty of Medicine were screened for C trachomatis and N gonorrhoeae by Abbott's ligase chain reaction (LCR) using either urethral swabs or first-void urine. N gonorrhoeae cultures were done on a subset of these patients. RESULTS: The study enrolled 813 men. All of the men denied condom use during their previous sexual exposures. The overall prevalence of C trachomatis, as determined by LCR, was 15.7%. Only 192 patients were screened for both organisms. N gonorrhoeae prevalence was 9.4%. There was no difference in the chlamydia prevalence between men who had contact with commercial sex workers (CSWs) and men who had no such contact (15.3% versus 17.2%). However, clients of foreign CSWs were more likely to have chlamydia than clients of registered Turkish CSWs. CONCLUSIONS: C trachomatis and N gonorrhoeae are commonly found in Turkish men with urethritis. The findings did not show more chlamydial infection among men who had contact with CSWs than among men who had no such contact. The failure to use condoms among these men must be addressed.

Sexually transmitted diseases (STDs) in the world.

Sexually transmitted diseases (STDs) represent a major public health problem in the world and the advent and increase of human immunodeficiency virus infection during the last decade has highlighted the importance of infections spread by the sexual route. The World Health Organization estimates that the global incidence in 1995 of new cases of selected curable STDs, which are gonorrhea, chlamydial infection, syphilis and trichomoniasis, was 333 million. Control programs for STDs must prevent the acquisition of STDs, their complications and sequelae and interrupt and reduce transmission.

Screening of sex workers in Turkey for Chlamydia trachomatis.

BACKGROUND: Increasing migration of sex workers across East European borders into Turkey has resulted in increased arrests of unregistered sex workers. There is concern regarding the prevalence of sexually transmitted diseases (STD) such as C. trachomatis in this unregulated group. OBJECTIVE: To measure the prevalence of C. trachomatis infection and selected related demographic and behavioral factors among registered and unregistered sex workers in Istanbul, Turkey. METHODS: In a cross-sectional study, the prevalence of C. trachomatis infection was studied by direct fluorescent antibody testing in 248 sex workers: 158 presented for routine medical screening and 90 who were arrested for lack of registration. All subjects were interviewed regarding demographic factors, sexual activity, and antibiotic and drug use. RESULTS: The overall C. trachomatis prevalence was 12.9% (12.0% registered and 14.4% unregistered). There was a strong association between never using condoms and chlamydia positivity (odds ratio 8.9 [95% confidence interval 3.7 to 21.6]). CONCLUSIONS: The high rate of C. trachomatis among all tested sex workers and the lack of regular condom use, particularly among unregistered sex workers, indicate that there is an urgent need for safe sex education and continued STD screening of unregistered sex workers.

PIP: Accelerating migration of unregistered sex workers from Eastern European countries to Turkey has produced alarm about the potential for increased sexually transmitted disease (STD) transmission. Licensed sex workers are routinely screened for STDs and given medical examinations. However, in Turkey, women with foreign citizenship are ineligible for licensing. This cross-sectional study measured the prevalence of Chlamydia trachomatis in 248 sex workers in Istanbul in 1993. 158 sex workers were enrolled when they presented for routine medical screening, while 90 (from Rumania) were arrested for lack of registration. The overall C. trachomatis prevalence, as detected by direct fluorescent antibody testing, was 12.9% (12.0% among registered and 14.4% among unregistered women); however, the fact that almost half these women reported recent self-prescribed antibiotic use suggests these rates are underestimates. When the analysis was restricted to the 84 women without recent antibiotic use, the C. trachomatis prevalence was 36.9%. There was a significant association between never using condoms and chlamydia infection (odds ratio, 8.9; 95% confidence interval, 3.7-21.6). These findings suggest a need for safe sex education and continued STD screening of sex workers in Turkey. Eligibility for registration, regardless of nationality, should be reconsidered for public health reasons.

Clinical algorithms for the screening of Chlamydia trachomatis in Turkish women.

OBJECTIVE: To test the diagnostic validity of clinical algorithms for the detection of Chlamydia trachomatis in an urban population of married women in Turkey. DESIGN: Cross-sectional population-based survey. SUBJECTS: A systematic sample of 867 women who reported the use of contraceptive methods. MAIN OUTCOME MEASURES: Sensitivity, specificity and positive predictive value of clinical algorithms for the diagnosis of C trachomatis. RESULTS: C trachomatis was diagnosed in 4.89% of the women. The WHO algorithm for use in settings where no vaginal examination could be performed had a sensitivity of 9% and a specificity of 96%. The corresponding figures for the WHO algorithm incorporating the findings of a speculum examination were 47% and 56% respectively. Algorithms incorporating symptoms or signs other than those suggested by the WHO did not yield satisfactory standards of validity. CONCLUSIONS: The findings of this study do not support the widespread introduction of the use of clinical decision models for screening of women for chlamydia infection in primary health care settings such as family planning or antenatal clinics. The large number of false positive results with the use of the clinical algorithms tested in this study would cause unnecessary costs to the health system and unnecessary interventions to the women treated.

Prevalence of hepatitis B virus infection in pregnant Turkish women and their families.

A total of 5,366 pregnant Turkish women were screened for hepatitis B surface antigen (HBsAg) and 225 (4.2%) of them were found to be positive. Hepatitis B e antigen (HBeAg) was detected in 6.2% of HBsAg-positive pregnant women. the overall prevalence of HBsAg and antibody to HBsAg (anti-HBs) among the spouses, previous children, mothers and first degree relatives of the HBsAg-positive pregnant women was 56%, 49%, 79% and 74% respectively. The prevalence of HBsAg is thus high in pregnant Turkish women with familial clustering of hepatitis B virus infection.

The treatment of non-gonococcal urethritis with single dose oral azithromycin.

In an uncontrolled study, the efficacy of azithromycin in the treatment of non-gonococcal urethritis was assessed in 41 male patients aged between 20 and 40 years with a mean age of 27 +/- 5 years. Clinical and microbiological diagnosis confirmed that 28 men were found positive for Chlamydia trachomatis, 10 for Ureaplasma urealyticum and three for both C. trachomatis and U. urealyticum. All patients received 1 g azithromycin orally (four 250 mg capsules). The length of time between the treatment and following visits were 7-10 days and 14-21 days for second and third visits, respectively. Complete eradication was achieved in 27 out of 41 patients. Of the remaining 14, six were found positive for C. trachomatis and were excluded as they did not return for the follow-up visit, one patient did not achieve complete eradication, one patient infected with both C. trachomatis and U. urealyticum failed to achieve complete eradication, and six patients infected with U. urealyticum failed to be completely cured. No adverse effects were reported in any patient. Single dose administration of 1 g azithromycin appears to be an effective and well-tolerated treatment for chlamydial urethritis and an advantage in terms of patient compliance.

In vitro activity of azithromycin (CP-62,993) against Chlamydia trachomatis and Chlamydia pneumoniae.

The in vitro susceptibilities of 49 strains of Chlamydia trachomatis and 3 strains of Chlamydia pneumoniae to azithromycin and tetracycline or doxycycline were determined. The MIC of azithromycin ranged from < or = 0.06 to 1.0 micrograms/ml, the MIC of tetracycline ranged from 0.03 to 0.12 micrograms/ml, and the MIC of doxycycline ranged from 0.015 to 0.06 micrograms/ml against C. trachomatis. The MIC ranges for C. pneumoniae were 0.12 to 0.25 micrograms/ml for azithromycin and 0.06 to 0.12 micrograms/ml for tetracycline. All minimal chlamydicidal concentrations were either equal to the MIC or one or two dilutions higher. No strains resistant to these antibiotics were detected. In vitro activity shows that azithromycin is highly active against C. trachomatis and C. pneumoniae.