In this study, the efficacy and safety of assisted reproduction techniques with the sperm-washing method and nested PCR assay were evaluated in HIV-1-discordant couples, as many HIV-1-positive people of reproductive age are getting married and wish to have children safely. Twenty-seven HIV-1-discordant couples (husband, positive; wife, negative) were enrolled in this study. The spermatozoa were separated from semen samples by density gradient centrifugation and the swim-up method. HIV-1 RNA and proviral DNA were checked using nested PCR with a detection limit of one copy before fertilization and before embryo transfer. Clinical outcomes were compared with those of matched control couples. Thirty-eight cycles of in vitro fertilization or intracytoplasmic sperm injection were performed in HIV-1-discordant couples, where the pregnancy rates per embryo transfer and per couple were 60.6 and 63.0%, respectively. These rates were significantly higher than those in control couples (P < 0.05). Furthermore, all of the females and babies remained HIV-1 negative throughout the study period. Our data strongly suggest that this technique will allow HIV-1-discordant couples to conceive more safely and effectively.

Two hundred and twenty-five fecal strains of Escherichia coli isolated from 109 non-hospitalized adults in 2006 were investigated for susceptibility to antibiotics and for the presence of integrons. High resistance rates in fecal strains of E. coli were observed for streptomycin (52.0%), ampicillin (50.2%), piperacillin (50.2%), trimethoprim/sulfamethoxazole (47.6%) and chloramphenicol (33.8%). Integrons were found in 31.5% (71/225) of the strains using an integrase gene PCR assay. Among 71 integrase-positive strains, 65 strains belonged to class 1 integrons, while the remainder belonged to class 2. Gene cassette patterns of class 1 integrons were further characterized by PCR and direct sequencing. Among those class 1 integrase-containing isolates, the integron cassette region was amplified by PCR in 40.0% (26 of 65) of isolates. Five different antimicrobial resistance gene cassette arrays were found in those isolates. These gene cassettes included those encoding resistance to trimethoprim (dfrV, dfrA7, dfrA12, dfrA17) and streptomycin (aadA1, aadA2, aadA5). Among those gene cassette arrays, dfrA12-orfF-aadA2 was found in 53.8% (14/26) of the isolates. These findings indicate that multidrug resistance of fecal flora is common in Taiwan and that integrons play an important role in resistance to trimethoprim and streptomycin in humans.

Medical treatment of pulmonary Mycobacterium avium complex (MAC) disease does not always provide curative effects and is frequently hampered by recurrence. This suggests the presence of a reservoir for MAC in the environment surrounding patients. We previously reported the recovery of MAC isolates from the residential bathrooms of outpatients. In the present study, to ascertain the colonizing sites and the possibility of an MAC reservoir in the bathrooms of patients, we tested the recovery and the genetic diversity of MAC isolates from 6 sites of specimens, including 2 additional sampling sites, inside the showerhead and the bathtub inlet, in the residential bathrooms of patients with pulmonary MAC disease. MAC isolates were recovered from 15 out of the 29 bathrooms (52%), including specimens from 14 bathtub inlets and 3 showerheads. Nearly half of these bathrooms (7/15) contained MAC strains that were identical or similar to their respective clinical isolates Additionally, in 5 out of 15 bathrooms, polyclonal colonization was revealed by pulsed-field gel electrophoresis. The results imply that colonization of MAC organisms in the bathrooms of MAC patients occurs predominantly in the bathtub inlets, and there is thus a risk of infection and/or reinfection for patients via use of the bathtub and other sites in the bathroom.

In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was established which can detect 10³ copies of viral RNA corresponding to approximately 5 fg of RNA. RT-LAMP with the Phil primer set designed according to the nucleotide sequences obtained from a Kyoto patient who contracted rabies in the Philippines was able to amplify all 16 street viral sequences derived from the Philippines. The specificity of RT-LAMP products was easily confirmed by digestion with RsaI restriction enzyme. The reaction of RT-LAMP could be completed within 1 h and could be conducted under isothermal conditions using a conventional water bath or heat blocks, indicating that RT-LAMP is ideal for the diagnosis of rabies in developing countries. Although further study is required to establish more universal RT-LAMP primers applicable to viruses from other regions or countries, the fast, easy, simple, sensitive and specific RT-LAMP method established here might be useful for rabies diagnosis and can facilitate studies of rabies epidemiology where rabies is enzootic, particularly in developing countries.

Group B Streptococcus (GBS) infection was studied in 49 patients collected at convenience (convenience sampling), excluding infants and women with genital tract- and pregnancy-related isolates, according to the availability of stocked isolates and easy accessibility to epidemiological data. The data were examined both prospectively and retrospectively from 2003-2005 at a tertiary-level multidisciplinary hospital in Kuala Lumpur, Malaysia. Skin and soft-tissue infections in 35 patients (71.4%) were the most common clinical presentation, while diabetes mellitus was the most common underlying condition (35 patients, 71.4%). All GBS isolates were sensitive to penicillin, and most isolates tested were sensitive to erythromycin (97.7%). Serotyping of 45 GBS isolates using a commercial serotyping kit revealed that the most common serotype was Ia (22.2%), followed by VI (17.8%), III and V (13.3% each). Others included Ib, II, IV, VIII, and VII; 13.3% were nontypeable. The findings of this pilot study are limited by the small sample size, the sampling method and the possibility that the cases are not wholly representative of the University Malaya Medical Centre population. Further studies from our hospital with larger numbers and using probabilistic sampling techniques are required to confirm the relatively high occurrence of serotype VI (the second most common serotype) in the population studied.

Persistent infection of arteries with organisms such as Chlamydia pneumoniae was previously found to contribute to the development of atherosclerosis. We investigate the presence of C. pneumoniae in atherosclerotic plaque by polymerase chain reaction and direct immunofluorescence assay, and we examine the correlation between clinical status and the presence of this bacterium in Iran. The study group consisted of 33 atherosclerotic plaque specimens from the arteries (26 coronary and 7 abdominal aorta) of patients who had undergone coronary artery bypass grafting surgery (CABG). The control group consisted of 31 specimens: 12 from biopsies of macroscopically healthy regions of the ascending aorta in patients who had undergone CABG and 19 autopsy specimens of normal coronary arteries. C. pneumoniae DNA and antigen were found in 6 (18%) and 7 (21%) of 33 endarterectomy specimens, respectively. C. pneumoniae was not detected in the control group by either method. The presence of C. pneumoniae in atherosclerotic plaques and its absence in healthy vessels supports the idea that C. pneumoniae may have a role in the development of atherosclerosis, especially in countries where infection is prevalent and where conventional risk factors fail to explain the exact reason for the high prevalence of atherosclerotic vascular disease.

We examined 29 isolates of Salmonella enterica subspecies enterica serovar Schwarzengrund from broiler chickens (n = 19) and retail chicken meats (n = 10) in Japan for antimicrobial susceptibility and pulsed-field gel electrophoresis (PFGE) profiling. All isolates exhibited resistance to both bicozamycin and sulfadimethoxine (minimum inhibitory concentration of both antimicrobial agents: ≥512 μg/ml). Nalidixic acid resistance was found in only one broiler chicken isolate. PFGE analysis showed that there were two genotypes among S. Schwarzengrund isolates. Isolates from 11 of 19 broiler chickens and from 6 of 10 retail chicken meats exhibited resistance to dihydrostreptomycin, kanamycin, oxytetracycline, bicozamycin, trimethoprim, and sulfadimethoxine, and had an identical PFGE pattern classified into a predominant genotype. Thus, our results indicate that genetically identical multidrug-resistant S. Schwarzengrund appeared to be disseminated among broiler chickens and retail chicken meats in Japan.

To evaluate the relationship between the incidence of legionellosis and Legionella concentrations in bathwater, we sent a questionnaire to 76 prefectural and municipal public health laboratories in Japan and found that 35 had encountered cases of legionellosis and had implemented investigations to determine the sources of the infections. Based on the results of the questionnaire, we were able to analyze various characteristics of the patients, of the facilities that were thought to be associated with the cases, and of the species and serogroups of the isolates and concentrations of Legionella. Ninety-six cases were included in this study. The median age was 67 years (range, 13-89 years). The most prevalent underlying medical condition among patients was diabetes, and the second most prevalent was high blood pressure. Concentrations of Legionella in bathwater ranged from 10 to 160,000 CFU/100 ml. Ten episodes were selected in which causative strains were found in the suspected source environment, and were then confirmed by pulsed-field gel electrophoresis analysis, enabling us to provide an estimated infectious concentration range of Legionella of 90 to 140,000 CFU/100 ml. It was thus suggested that the current Japanese regulatory safety level for Legionella in bathwater, which is set below the detection limit of culture techniques (10 CFU/100 ml), should be appropriate to prevent bathwater-associated legionellosis. In tandem with the above-mentioned research, a review of literature concerning bathwater-associated legionellosis and typical cases was undertaken.

Emphysematous pyelonephritis (EPN) is a rare but severe infectious disease. This disease sometimes presents bilaterally, making it difficult to cure. Diabetes mellitus is a common cofactor in this disease. Drainage or nephrectomy, often combined with antibiotics, is ordinarily used for treatment. To our knowledge, only 8 cases of bilateral EPN cured by antibiotics alone have been reported. We report the case of an 86-year-old woman with bilateral EPN cured by antibiotic therapy alone, thus avoiding surgery or drainage in a frail elderly patient.

In 2007 eight epidemic clusters (more than 15 cases in each) and other sporadic cases of measles occurred in Hokkaido district, Japan. A total of 850 cases were identified. Approximately half of them were ≥15 years of age, resulting in a huge public health problem in the community associated with school closings, and 31% of the cases reported that they already had a history of vaccination. Of 28 isolates of the measles virus detected, all were identified as genotype D5, identical to the type isolated in other areas of Japan, suggesting that a highly homologous measles virus circulated in Japan. The occurrence pattern of measles patients and molecular epidemiology indicated that the measles virus that spread in Hokkaido district might not be indigenous.

Most first-line anti-tuberculosis drugs have less in vitro activity against atypical mycobacteria. Loop-mediated isothermal amplification (LAMP) was used for the rapid diagnosis of mycobacterial species. The sensitivity of LAMP was 96.1% (49/51) in smear-positive and culture-positive sputum samples and 85.0% (17/20) in smear-negative and culture-positive samples. Of the 77 total LAMP-positive samples, 75 (97.4%) were identified as Mycobacterium tuberculosis and 2 (2.6%) as M. intracellulare. One of the M. intracellulare-infected cases was identified in a patient with suspected mycobacteriosis and another was found in a follow-up patient.

The nucleotide sequences of the inner repeats of the repetitive sequence (RPS), termed ALTs, of Candida albicans and its related species C. albicans var. stellatoidea and C. dubliniensis, were analyzed. ALT sequences were grouped into 4 types for C. albicans (Aa, Ab, Ac and Ad) and C. albicans var. stellatoidea (Sa1, Sa2, Sb, Sc and Sd), and 3 types for C. dubliniensis (Da, Db and Dc). In addition to the primer set P-II (specific to RPS), 2 primer sets (AS-I and AiR-I) specific to the nucleotide sequences of C. albicans ALT were designed and tested for their potential for RPS-based identification/genotyping of C. albicans. PCRs using AS-I and AiR-I clearly distinguished C. albicans from both C. albicans var. stellatoidea and C. dubliniensis. Furthermore, the strains of C. albicans that showed similar electrophoretic patterns in the PCR using P-II were discriminated at the subtype level. These results indicate that the PCRs using RPS- and ALT-specific primer sets are useful as simple and rapid systems for the specific identification and genotyping of C. albicans.

Serum specimens were collected from 125 pigs on Miyako Island, 112 pigs on Ishigaki Island, and 42 pigs on Kume Island from 2005 to 2007, and 54 pigs on Yonaguni Island from 2006 to 2007. Their sera were tested for Japanese encephalitis virus (JEV) antibody by hemagglutination inhibition (HI) assay. Five serum samples (4.5%) from Ishigaki Island were positive for HI antibody, and 4 of the 5 samples were positive for 2-mercaptoethanol-sensitive antibody (IgM Ab). All samples from Miyako, Kume, and Yonaguni Islands were negative for HI antibody. Our results indicate that JEV transmission activity was extremely low on Miyako, Ishigaki, Kume, and Yonaguni Islands. The JEV genome (JEV-RNA) was detected from the sera of one pig on Ishigaki Island. The partial gene of the E region (151 nt) was analyzed phylogenetically. The analysis showed that the new JEV-RNA belonged to genotype 3 and was closely related to JEV strains isolated in Taiwan from 1985 to 1996. It was suggested that JEV previously introduced from Taiwan had been maintained on Ishigaki Island.

The ¹³C-urea breath test (UBT) is widely used to examine the results of eradication therapy for Helicobacter pylori. We examined whether a stool antigen test can be used as efficiently as UBT. (i) Ninety-four patients infected with H. pylori underwent eradication therapy. Six or eight weeks after the completion of treatment, the results were evaluated by UBT and a stool antigen test (TPAg). In 77 out of 78 patients who had negative UBT results, the TPAg results were also negative. Among the 16 UBT-positive patients, 12 were also positive by TPAg. Agreement of UBT and TPAg was 94.7%. (ii) Twenty-two patients with peptic ulcers in the active stage also received eradication therapy followed by proton pump inhibitor (PPI) administration. TPAg and UBT were performed to examine the results of eradication therapy at the end of PPI administration and at least 7 days after its discontinuation. Of the 22 patients taking PPIs, TPAg evaluated the results of eradication therapy accurately in 21 patients. TPAg appears to be an accurate test for evaluating the results of H. pylori eradication therapy, and to be as efficient as ¹³C-UBT. Use of the stool antigen test should be considered as an alternative, particularly in patients who have to take PPIs in order to avoid the risk of peptic ulcers.

Methicillin-resistant Staphylococcus aureus (MRSA) is a problem within healthcare organizations and in the community. The aims of this study were to identify the prevalence of S. aureus in the anterior nares of surgical unit staff, to analyse their antibiogram with special reference to methicillin resistance, and to compare the isolates among surgical unit staff and in relation to the wards where they worked. Sterile swabs were used to collect the samples from the anterior nares of 100 healthcare workers working in 5 surgical wards who satisfied rigid inclusion and exclusion criteria. Standard procedures were followed for isolation, identification, and antibiotic sensitivity testing. S. aureus carrier status was observed in 13 individuals, of whom 2 (15.4%) were resistant to methicillin. All the isolates of S. aureus were multidrug-resistant but sensitive to vancomycin and bacitracin. One of the 13 was resistant to linezolid. Sixty-three of the staff were carriers of coagulase-negative Staphylococcus. The presence of methicillin resistance may cause problems in hospital infection control programs and may indicate emerging issues. This study suggests the need for periodic screening of hospital personnel in order to monitor trends and take steps to treat carriers.

In this study, the patterns of resistance to 10 antibiotics by 730 Vibrio cholerae non-O1 and non-O139 species isolated from both environmental and seafood samples were investigated. Susceptibility to different antimicrobial agents was assessed by the disc diffusion technique. The frequencies of resistance to 10 antimicrobial agents—ampicillin, chloramphenicol, bacitracin, erythromycin, gentamycin, streptomycin, oxytetracycline, vancomycin, penicillin, and neomycin—were 88, 46, 8, 64, 13, 85, 18, 21, 84, and 18%, respectively. About 10-20% of the studied strains showed a 3-5 multiple drug resistance pattern.

Surveillance is critical for both early detection and a timely public health response to infectious diseases. Electronic information systems have been widely adopted by pharmacies in Japan. The aim of this study is to evaluate the feasibility of using prescription data for influenza surveillance in Japan to facilitate the development of a daily reporting system. This is a retrospective study using questionnaires mailed to pharmacies in Sakai City, Osaka, Japan in October 2007. The total number of prescriptions for oseltamivir and zanamivir and the number of influenza cases reported by sentinel surveillance in Sakai during the investigation period showed excellent correlation, with a correlation coefficient of 0.954. Further analysis showed that the data from as few as eight pharmacies result in a correlation coefficient of 0.9. These results demonstrate the feasibility of such a system and that pharmacy prescription data are a very useful indicator of sentinel surveillance for influenza.

An immunocompetent 8-year-old boy with cytomegalovirus (CMV)-associated transient protein-losing enteropathy (PLE) is described. Colonoscopic examination revealed lymphoid hyperplasia of the terminal ileum. Histological examination of the biopsied specimens showed marked dilation of the lymphatic vessels. Primary CMV infection was demonstrated by serological test and polymerase chain reaction. The child had complete resolution of the disease without antiviral treatment. The present case suggests the etiologic role of CMV infection in PLE resulting from intestinal lymphangiectasia in childhood.

We encountered five cases of pulmonary Mycobacterium kansasii (MK) disease for which the QuantiFERON-TB Gold (QFT-2G) test was useful as a supportive diagnostic method. Because none of these patients could expectorate sputum, we ultimately confirmed the clinical diagnosis of pulmonary MK disease using 6- to 8-week cultures of bronchoscopic specimens (bronchoalveolar lavage fluid) according to the guidelines for nontuberculous mycobacterial disease due to American Thoracic Society/Infectious Diseases Society of America. The tuberculin skin test showed positive responses in all cases, and the QFT-2G test showed positive response for ESAT-6 only or CFP-10 only antigens. After it was proved that the result of Mycobacterium tuberculosis (MTB)-nucleic acid amplification was negative and the result of the QFT-2G test was positive, we started treatment (isoniazid, rifampicin and ethambutol) for suspected pulmonary MK disease based on clinical findings in all cases. The clinical effect was good and the interferon-γ response to MTB-specific antigen decreased with treatment for pulmonary MK disease. The QFT-2G test may be useful as a rapid supportive diagnostic method for pulmonary infection due to MK possessing the same MTB-specific antigen.