The simplest, cheapest, and fastest diagnostic method for tuberculosis (TB) is the detection of acid-fast bacilli (AFB) by microscopy. The algorithm advised for the diagnosis of TB recommends examination of three consecutive sputum specimens from TB suspects for the presence of AFB. In the present study, we evaluated the contribution of each specimen to the final detection of TB suspect patients with culture-proven disease. The collection and analysis of retrospective data on patients with culture-proven pulmonary TB, from June 2002 to August 2006 at Dokuz Eylul University Hospital, Turkey, have enabled us to assess the value of examining two sputum specimens in diagnosing this disease. AFB were detected from one or more sputum specimens with direct microscopy in 42% of the cases. An analysis of results of smear examination showed that 97% of AFB were detected from the first specimen and only 3% were obtained from the second smear. The third specimen did not have any additional diagnostic value for the detection of AFB by microscopy. As a conclusion the present study shows that examining two sputum smears is sufficient for the early detection of AFB in our laboratory.

Data on various etiologic agents causing diarrhea in human immunodeficiency virus type-1 (HIV-1) infected individuals are sparse in Delhi, India. The present study was undertaken to identify various causative agents, the role of associated risk factors and immune status. A case-control study was conducted among 75 HIV-1 infected individuals, 50 with and 25 without diarrheal infection. Fecal samples were screened for coccidian parasites, enteric protozoa, and helminthes by using various staining techniques. The CD4⁺ T-lymphocyte count was estimated. Enteric parasites were identified among 62.7% individuals, of which Cryptosporidium emerged as the single largest pathogen predominant among 33% of the individuals (P < 0.025). Other parasites diagnosed that were significantly associated with diarrhea were Giardia lamblia (13.3%), microsporidia (6.7%), and Isospora belli (2.7%). Chronic infected diarrheal cases were found to have polyparasitic infections. The mean CD4⁺ cell count was found to be lower among the diarrheal cases when compared with the non-diarrheal cases (mean, 141 cells/mm³ versus 390 cells/mm³). Similarly, among diarrheal individuals, the chronic diarrheal cases had a comparatively lower CD4⁺ cell count than the acute cases (mean, 123 cells/mm³ versus 265 cells/mm³). Risk factors found significant during multivariate analysis were: residence in a slum, exposure to pets and animals, use of public toilets, and practice of unsafe homosexual activity. Enteric coccidian parasites were identified as significant agents associated with diarrhea, especially among those with improper hygiene, multiple infections and a lower CD4⁺ cell count. Thus, this study emphasizes the need for routine screening of enteric parasites as well as education about practicing personal hygiene and taking timely and appropriate prophylactic measures.

The purpose of this study was to type the Pseudomonas aeruginosa and Stenotrophomonas maltophilia isolates recovered from cystic fibrosis (CF) patients by random amplified polymorphic DNA (RAPD)-PCR and to determine the antibiotic susceptibility of these strains. P. aeruginosa (n = 49), and S. maltophilia (n = 11) isolates which had been recovered from 16 and 8 patients, respectively, during a 1-year period were investigated. Three primers were used for RAPD-PCR typing. Antibiotic susceptibility testing of all isolates was performed by the disc diffusion method. RAPD-PCR analysis revealed 21 (P. aeruginosa) and 9 (S. maltophilia) different genotypes. According to the antimicrobial susceptibility results, the P. aeruginosa and S. maltophilia strains were cumulated into 24 and 11 groups, respectively. The CF patients were colonized or infected with P. aeruginosa strains of single or sometimes multiple genotypes which remained stable over several months. Our results also revealed that cross-colonization might be possible among the patients who are followed up at the same center. Piperacillin-tazobactam for P. aeruginosa and trimethoprim-sulfamethoxazole for S. maltophilia were found to be the most active antibiotics according to our results.

Few studies have investigated the risk factors for nosocomial infections developed in neurology intensive care units (ICUs). In this study, the risk factors for ICU-acquired infections in patients with cerebral hemorrhage and cerebral infarct who were treated for more than 24 h at the Ankara Training and Research Hospital were prospectively evaluated over a study period of 14 months. Of 171 patients included in the study, 71 (41.5%) were found to have acquired 163 infections in the ICU unit throughout 1,867 patient days. The rate of infection per 100 patients admitted was 95.3, and per 1,000 patient days, 87.3. The most common nosocomial infections were urinary tract infection (42.9%), pneumonia (27%) and primary bacteremia (19%). Multivariate logistic regression analysis revealed age ≥ 70 (P < 0.05), the presence of a central venous catheter (P = 0.004), and parenteral nutrition (P = 0.02) as ICU-acquired infection risk factors. The presence of infection on admission was identified as a factor decreasing the risk of ICU-acquired infection (P < 0.001). The high infection rates found in this study may be due to lack of full compliance to infection control measures. In conclusion, each type of ICU has its own epidemiological findings for nosocomial infections and thus needs to determine the risk factors using periodical surveillance studies to guide control measures.

In order to assess the effect of the neonatal tetanus elimination program in Turkey, tetanus antibody prevalence among women of childbearing age from three selected provinces was evaluated in relation to vaccination doses of the single-type tetanus vaccine. A combined method of in-house enzyme-linked immunosorbent assay and particle agglutination test was used to determine tetanus antibody titers. Among 205 women aged 20 - 39 years, the tetanus antibody level was higher in women with 1 - 3 children than those without children. The geometric mean of the log antibody titer was increased proportionally with a slope of 0.405 ± 0.174 per dose between 0 and 3 doses (P > 0.05). However, the proportion of 20 - 39-year-old women with the protective antibody in the provinces ranged from 54.8 to 86.6%. Diyarbakir had the lowest immunity with a larger number of children in the household, and a lower educational level. The results of our serological study demonstrated that the neonatal tetanus elimination program in Turkey is effectively promoting immunity against tetanus in pregnant women. However, the study also revealed that the tetanus immunity among women of childbearing age was still insufficient. Intensive implementation of the supplemental immunization activities and encouraging vaccinations through neonatal care services will improve the situation.

During endemic infections, the sensitivity of diagnostic tests and rapid diagnosis of the respiratory tract pathogens is particularly important. Utilization of just one diagnostic technique, such as serological tests or polymerase chain reaction (PCR)-based detection methods, during outbreaks of lower respiratory tract infections (LRI) can result in some of the patients being missed. In this study we aimed to investigate the etiology of LRI in military recruits in Izmir, Turkey, among whom several pneumonia cases have been reported and 47 patients have been hospitalized. Nasopharyngeal swabs were used for PCR analysis of Chlamydophila pneumoniae, Mycoplasma pneumoniae and Legionella spp. Serum samples were collected in the acute and convalescent phase of infection for C. pneumoniae and M. pneumoniae. Thirty-nine patients were diagnosed with C. pneumoniae infection by PCR and/or serology. Diagnoses were established by PCR in the acute phase of infection in 40.4% of the group. Based on the results of these studies, PCR is a useful method for early detection and identification of C. pneumoniae-related LRI outbreaks. However, this technique is not sufficient to detect all positive cases per se. After effective therapy and introduction of appropriate infection control measures, the outbreak ceased without mortality. This is the first closed-community C. pneumoniae outbreak report from Turkey.

The aim of this study was to compare the efficacy of the tuberculin skin test (TST) and the quantiferon test (QFT) for detecting latent tuberculosis infection (LTBI) in health care workers (HCWs). Seventy-six participants who were working in Duzce University Hospital, where tuberculosis patients were being treated, were included in the study. TST was performed according to the Mantoux technique. QFT was performed in accordance with the manufacturer’s instructions. A positive TST result was defined as an induration diameter of ≥15 mm. TSTs were positive in 41 of 76 participants (53.9%) and QFT was positive in 65 of 76 participants (85.5%). There was a significant difference between the numbers of QFT-positive and TST-positive cases (P = 0.02). When the induration diameter of TST was ≥20 mm, QFT positivity was 100%. Multivariate analysis revealed that there was a significant correlation between the percentage of patients with QFT positivity and the induration diameter of TST (P = 0.009). QFT thus seems to be more effective for LTBI diagnosis than TST. However, large-scale trials including quantitative measurement of QFT in subgroups taking into account the division where HCWs are employed and the different results of TST might clarify the usefulness of QFT in LTBI diagnosis.

The demand for rapid and simple development of a vaccine against a newly emerging infectious disease is increasing worldwide. We previously revealed that UV-inactivated severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) virions (UV-V) elicited high levels of humoral immunity and a weak Th0 response in mice immunized subcutaneously. To ensure the safety of such a whole inactivated SARS-CoV vaccine, we additionally treated the UV-V vaccine with formalin, resulting in the UV-F-V vaccine. Analysis of the immunogenicity of the UV-FV+alum vaccine in mice revealed that it generated comparable neutralizing serum anti-SARS-CoV IgG antibody levels as the UV-V+alum vaccine. Moreover, both vaccines induced similar frequencies of anti-SARS-CoV IgG antibody-producing cells in bone marrow. Interestingly, the UV-F-V vaccine induced fewer IgG_2a subtype antibodies and higher interleukin-4 production in vaccinated mice than did UV-V. Thus, UV-F-V imposes a Th2-type bias on the immune response, unlike UV-V. We propose here that doubly-inactivated SARS-CoV virions by UV and formalin constitute a safe vaccine that may effectively induce neutralizing antibodies in humans.

Highly active antiretroviral therapy (HAART) can suppress human immunodeficiency virus type 1 (HIV-1) replication and plasma HIV-1 to below detectable levels. However, HAART becomes ineffective when drug-resistant viruses emerge during HAART. Monitoring drug-resistance mutations in viruses is necessary for selecting new drugs or therapies effective at inhibiting such HIV-1 variants. Most laboratories in Japan perform the tests using in-house protocols. However, the quality of these tests has never been assessed. Our study assessing the accuracy and reliability of HIV-1 genotypic drug-resistance testing in 15 laboratories in Japan revealed that the quality was very high (97.3% accurate). The errors, though rare, were caused by human errors, poor electropherograms, and the use of inadequate primers. Here, we propose troubleshooting procedures to improve testing accuracy and reliability in Japan.

Japanese encephalitis virus (JEV) formed plaques in mosquito C6/36 cell layers after adsorption on the cell surface and exposure to pH values lower than 6.2. The number of plaques decreased within pH ranges from 7.4 to 6.4, but increased within pH ranges from 6.2 to 5.8. Plaque formation was prevented by treatment of the virus with a JEV-neutralizing monoclonal antibody, 503, after virus adsorption. Plaque formation was not affected by pretreatment with a specific V-ATPase inhibitor, bafilomycin A1. The results indicate that JEV successfully fused with the C6/36 cell membrane under acidic conditions below pH 6.2, which in turn led to plaque formation in C6/36 cell layers. These results suggest that productive JEV infection occurs at the C6/36 cell surface via the fusion between JEV and the cell membrane.

The aim of this study was to isolate, identify and compare the pathogenic bacteria prevalent on the overcoats of doctors, residents and students from medical and surgical wards, to determine their antimicrobial sensitivity, compare them with isolates from pus collected over the same period, and ultimately make recommendations. Using standard procedures, bacteria were isolated and identified from the hem and pocket mouths of the overcoats of 80 medical personnel, and drug sensitivity tests were carried out. Of the samples from the overcoats, 95% (n = 152) were positive for bacterial isolates like Pseudomonas aeruginosa, Klebsiella sp., Escherichia coli, non-fermenting Gram-negative bacteria, Staphylococcus aureus, etc. Six swabs showed double isolates. There was a significant (P = 0.014) association with the category of the participants (30/34 from doctors, 44/48 from residents and 78/78 from students). The isolates were significantly (P < 0.001) more prevalent on overcoats from surgical wards (n = 98, 100%) than on those from medical wards (n = 54, 87%). The pathogens from medical overcoats and those from pus samples were both multidrug resistant, though they were not similar. Hence overcoats may be a transmitting agent for bacterial pathogens. Doctors should be aware of the proper usage and frequency of laundering of overcoats.

Antimicrobial resistance to nine anti-pseudomonal agents (azteronam, ceftazidime, cefepime, piperacillin/tazobactam, imipenem, meropenem, ciprofloxacin, amikacin and gentamicin), the magnitude of multidrug resistance, associated underlying conditions, and mortality among patients with Pseudomonas aeruginosa isolates from King Khalid University Hospital, Riyadh, Saudi Arabia from 2001 to 2005 were determined. The results showed that antimicrobial resistance among P. aeruginosa is gradually increasing for most anti-pseudomonal agents, particularly aztreonam, ceftazidime, piperacillin/tazobactam and imipenem. There were 19 (3%) and 12 (2%) multidrug-resistant (MDR) P. aeruginosa patients in 2004 and 2005, respectively, and MDR P. aeruginosa was more commonly found in non-intensive care unit (ICU) patients. Most MDR isolates were from surgical and diabetic patients. The mortality rate was higher among ICU patients.

Neonatal sepsis is a life-threatening emergency, and any delay in treatment may cause death. Because of the importance of the problem in Iran, the aim of this retrospective study was to determine the etiological agents of neonatal septicemia, and the prevalence and epidemiology of Klebsiella bacteremia in the neonatal wards. Two hundred and ten cases of neonatal sepsis occurred during the study period. The most common organism was coagulase-negative staphylococci. Gram-negative organisms were isolated in 66 cases (31.43%), and the most common Gram-negative organism causing neonatal sepsis was Klebsiella pneumoniae. The mortality rate due to Gram-negative bacteria including K. pneumoniae was higher than that due to other bacteria. The distribution of the main pathogens is different in the Azerbaijan state, northwest of Iran, and K. pneumoniae is predominant, but Streptococcus agalactiae plays a relatively minor role in the etiology of sepsis during the first month of life.

Acute osteomyelitis (AO) in neonates, although rare, represents a diagnostic and therapeutic challenge. A high index of suspicion is necessary to make an early diagnosis, and the observation of clinical signs is crucial. The increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) is an emerging problem in pediatrics. In neonates, MRSA infections can cause a wide spectrum of diseases including bone and joint infections. We report two cases of AO in full-term neonates, with no risk factors, due to MRSA.

We conducted a survey of diarrhea stool samples in which no virulent agents had previously been detected at clinical laboratories. DNA extracted directly and purified from the diarrhea stool was tested for bacterial pathogenic genes by polymerase chain reaction. The test results for 85 specimens were as follows: one sample was positive for lt, ipaH, and eae; two were positive for aggR; and eight were positive for astA. Inoculation with the stool specimens led to the isolation of a strain of Escherichia coli possessing eae, three strains of E. coli possessing astA, and a strain of Klebsiella possessing astA.

We present a case of cytomegalovirus (CMV)-induced pseudotumor of the gastric antrum. Although affliction of the entire gastrointestinal tract with CMV has been described, localization to the stomach and especially the gastric antrum is rare. Kaposi’s sarcoma and non-Hodgkin’s lymphoma are recognized causes of bowel thickening and obstruction in patients with AIDS, but CMV is an extremely rare cause, with only four cases of CMV-induced pseudotumor reported in the English literature. As the duration of opportunistic infections and length of survival of patients with AIDS increase, CMV pseudotumors are not likely to remain unique. This mass lesion should be included in the differential diagnosis of AIDS patients, along with Kaposi’s sarcoma and non-Hodgkin’s lymphoma.

We have developed a combinatorial polymerase chain reaction (PCR) procedure to identify four major species of the genus Brucella simultaneously. Four pairs of primers targeting the genes encoding a cell surface protein (BCSP31) and outer membrane proteins (omp2b, omp2a and omp31) were prepared. PCR using these primers gave rise to specific patterns of amplification for each Brucella spp. examined in this study. B. abortus could be identified when fragments of BCSP31 and omp2b/2a were amplified by B. abortus-specific primers. B. melitensis could be identified by the amplification of fragments of BCSP31, omp2b/2a and omp31 using pair of primers B4/B5, JRF/JPR-ab and omp31. Identification of B. canis could be achieved when the amplicons of omp2b/2a were detected by B. canis-specific primers, as could the identification of BCSP31 and omp31. If specific amplifications occurred using all pairs of primers, the strain was identified as B. suis. Combinatorial PCR reported here thus appeared to be an ideal method of identifying Brucella spp., the causative pathogen of human brucellosis.

Most mycobacterial infections are still caused by Mycobacterium tuberculosis complex (MTC) strains; however, infections by non-tuberculous mycobacteria (NTM) are increasing, particularly among immunocompromised patients. Conventional species-specific identification and proper patient management are delayed due to the slow-growing nature of mycobacteria. We have developed a multiplex PCR (mPCR) targeting the oxyR-ahpC intergenic region and rpoB gene for direct detection and differentiation of clinical isolates as MTC or NTM in primary culture. Two amplicons of 473 bp and 235 bp from MTC members and a single amplicon of 136 bp from NTM are expected. The mPCR was developed using several mycobacterial species and was evaluated by testing extracted DNA from liquid cultures, flagged as positive for bacterial growth, of 100 consecutive mycobacterial isolates. The results were validated by DNA sequencing of the species-specific 16S-23S internal transcribed spacer (ITS) region. The mPCR with template DNA from reference Mycobacterium spp. yielded the expected amplicons. When 100 consecutive clinical isolates of Mycobacterium spp. were tested, 92 strains yielded MTC member-specific amplicons, and DNA sequences from 10 randomly selected isolates matched completely with the ITS sequence from M. tuberculosis. Eight isolates were identified as NTM, and DNA sequencing of the ITS region confirmed the NTM status of each of these isolates. The mPCR developed in this study allowed rapid detection and differentiation of primary cultures as MTC or NTM, thus helping in timely institution of specific therapy.