We evaluated performance of Abbott RealTime CT/NG assay (real-time PCR, Abbott Japan) for detect Chlamydia trachomatis and Neisseria gonorrhoeae by real-time PCR in 88 female patients with cervicitis symptoms seen at gynecological clinics and 100 male patients with urethritis symptoms seen at urological or dermatology clinics in Kitakyushu, Japan. Endocervical swab and first-voided urine (FVU) specimens were then collected from women and FVU specimens from men. Detection rates of C. trachomatis and N. gonorrhoeae by real-time PCR in the 3 types of specimens were compared to those by ProbeTec ET assay (ProbeTec, BD Diagnostic System). The overall positive concordance between real-time PCR and ProbTec were 97.1％ (66/68)for C. trachomatis and 100％ (33/33) for N. gonorrhoeae, C. trachomatis detection yielded 3 discordant results in endocervical specimens and 1 discordant result in male FVU by real-time PCR and ProbTec. Three of 4 reexamined using Aptime Combo 2 Assay (Fuji Rebio Inc.) were positive for C. trachomatis. Endocervical swab and FVU specimen results for C. trachomatis were discordant in 3 cases in real-time PCR and 4 in ProbeTec. Subjects with 2 or more positive endocervical awab results in female or male FVU specimens were assumed to be “true positive” for C. trachomatis. The sensitivities of real-time PCR for detecting C. trachomatis was 94.4％ in endocervical swabs, 77.8％ in female FVU and 97.4％ in the male FVU. The sensitivities for real-time PCR for detecting N. gonorrhoeae was 100％in all 3 specimen types. Abbott RealTime CT/NG assay was useful for detecting C. trachomatis using endocervical swabs or male FVU specimens and for detecting N. gonorrhoeae using endocervical swabs and all FVU specimens.

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Object： Detecting of disease spread is an important task of public health and medical staff, especially in pandemics such as A/H1N1 flu (2009). This requires daily observation and estimation of the infected population. The fully automated real-time pharmacy survey we developed collects information electronically at pharmaceutical prescription. We used the data to analyze the pandemic A/H1N1 flu spread (2009) and to determine the systemʼs and capability in estimating the infected population. Method： Automatic collection of prescription information on anti influenza virus drugs from 3,959 pharmacies provided the basis for calculating the number of influenza sufferers and determining shape of the epidemic curve compared to that of official influenza sentinel surveys and mandatory reports of A/H1N1 (2009) patients. We also compared infection estimates from the pharmacy survey to those of official sentinel survey and a one-week survey of all hospitals and clinics in Gifu prefecture not reported in sentinel. Results： Fully automated real-time pharmacy surveillance began on April 20, 2009, and provided feedback at 07 : 00 daily. It estimated the infected population at 22,708 when official sentinel surveillance recorded an average of 0.99 influenza visits per week in epidemic week 32 when publicly announced that the pandemic had began in Japan. By the end of March, epidemic week 12 in 2010, infected-population estimates totaled 9,234,289, and peaked on November 24 at 234,519 in one day. All A/H1N1 (2009) sufferers reported mandatorily until mid-July numbered 25,526.The pharmacy survey indicated that there were influenza nationalwide by the time the very first outbreak emerged in the Kansai (western Japan) area. The correlation coefficient for the pharmacy and official sentinel survey was 0.992 nationwide, exceeding 0.95 in which only 33 of Japanʼs 47 prefectures were counted. The estimated infected population in the pharmacy survey was half of that of the official sentinel survey. The pharmacy survey yielded almost the same number as the complete survey in Gifu prefecture, however. Discussion： Fully automated real-time pharmacy surveys are useful in long-term observation e.g. detection of rapid emergence, identifying the peak, and careful monitoring of reemergence. It was demonstrated as the leading indicator for the official sentinel surveillance because of high correlation among them. Information collected daily is very useful in early detection and estimating the affected population. The survey consistently uses the same estimation criterion and operates automatically and routinely, facilitating the comparison of the latest and past results. The pharmacy survey indicated that official sentinel survey estimates overestimate actual cases and thus require modification to ensure accuracy. The pharmacy survey thus appears to be very valuable as a tool in measuring for the second wave of A/H1N1 (2009) or whatever the next pandemic may be. It can, of course, be applied to diseases other than influenza, e.g. varicella, by following antivaricellazostervirus prescriptions and antibiotic drugs.

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Group A rotavirus G-serotyping by polymerase chain reaction (PCR) using university hospital subject samples in September 2003 to August 2004, September 2004 to August 2005, September 2005 to August 2006, and September 2006 to August 2007 showed the most common serotypes G1 and G3, detected in 27 and 33 subjects, compared to 4 subjects in whom serotype G4 was detected. Between 2003 and 2004, serotypes G1 accounted for 50％ and G3 for 38％, contrasting with serotype G3 at 79％ between 2004 and 2005, serotype G1 at 91％ between 2005 and 2006, and serotype G1 and G3 at 37％ and 63％between 2006 and 2007, respectively. Serotypes G2 and G9 were not detected at all during any of our time periods. No correlation was seen between subject age and G serotype, although subjects younger than two years old accounted for 73％ of subjects. This infection caused combined fever, diarrhea, and vomiting in 48％ of subjects but showed no correlation with G serotype. These findings under-score the importance of G-serotyping in understanding rotavirus infection epidemiology at different times and indifferent locales.

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The presence in serum of the Hepatitis B surface antigen (HBsAg), the outer envelope of the hepatitis B virus (HBV), indicates viral infection, used in laboratory tests to confirm this. We report a case of discrepancy among HBsAg test results detected between measurements in a subject with HB infection. Gene analysis demonstrated several S region gene mutations, not detected previously. We tested 12 measurements e.g., EIA, CLIA, CLEIA, F-EIA, MAT, and IC for whether they could detect our subjectʼs HBsAg and found that it was not recognized by a method using only a single monoclonal antibody to detect HBsAg in two detection processes, in contrast to the 11 other measurements, which used two different antibodies. This case shows that amino acid substitution may cause a false negative result for HBsAg. Gene mutations known to occur in HBV, should thus trigger an awareness of the need to keep in mind that false negative results can happen in case such as ours.

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The minimum inhibitory concentation (MIC) and minimum bactericidal concentration (MBC) of 8 parental antibiotics were determined using 21 strains of Haemophilus influenzae from 21 children with invasive H. influenzae infection in 2006-2009. Children were from 4 months to 12 years, with 18 (85.7％) under 1 year old. Diagnosis involved meningitis or pneumonia in 7 (33.3％) ; epiglottitis, phlegmon, or arthritis in 2 each (9.5％), and occult bacteremia in 1 (4.8％). The capsule type was b (Hib) in 19 (90.5％). Classified by penicillin binding protein (PBP) gene mutation, strains were classified into (i) 6 β-lactamase negative ampicillin resistant (BLNAR) (28.6％), (ii) 5 low-BLNAR (23.8％), (iii) 4 β-lactamase positive clavulanic acid/amoxicillin resistant-II (19.0％), (iv) 2 β-lactamase positive ampicillin resistant (9.5％), and (v) 4 β-lactamase negative ampicillin susceptible (19.0％). Compared to a 2003-2005 survey, the number of sensitive strains had decreased significantly (p＜0.05). MIC₉₀/MBC₉₀ of ampicillin was 64/＞128μg/mL, piperacillin 16/＞128μg/mL, cefotaxime 0.5/0.5μg/mL, ceftriaxone 0.12/0.12μg/mL, panipenem 0.5/0.5μg/mL, meropenem 0.12/0.12μg/mL, doripenem 0.25/0.25μg/mL, and chloramphenicil 1/2μg/mL against H. influenzae.

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Luminex xTAG Respiratory Viral Panel FAST (RVP FAST) assay detects 17 human respiratory virus strains per measurement. Studying RVP FAST efficacy in detecting respiratory viruses in 67 aspirate samples from the nasal cavities of children with acute respiratory infection, we compared RVP FAST results to those of conventional nucleic acid amplification tests (NAT), e.g., real-time PCR, targeting 8 strains. RVP FAST assay detected 13 strains (98 isolates) in 59 of 67 samples. Of these, 8 -influenza virus (Inf.V) -AH1, Inf. V-AH3, novel Inf.V-AH1, and Inf.V-B, and adenovirus, RS virus, metapneumovirus, and bocavirus- were compared to NAT results. RVP FAST showed higher sensitivity (83.3-100％) and specificity (98.2-100％) than NAT. RVP FAST also detected coronavirus (CoV) 229E, OC43, NL63, and HKU1 from 10 virus strain samples and enterovirus and/or rhinovirus from 35. RVP FAST assay thus comprehensively detects clinically important viruses in a single measurement, making RVP FAST assay useful in detecting causative respiratory tract viruses.

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We discuss the efficacy 3 pandemic influenza, measures planned against an anticipated outbreak. First was an exclusive influenza outpatient clinic. Second was a medical call center for febrile illness subjects needing with fever clinic recommendation. The last was isolation. Before the outbreak, we had thought that all confirmed or suspected new influenza case should be quarantined. May 2009 brought the first A1/H1 pandemic influenza outbreak to Kobe, Japan. After the first infection announcement, call center and fever clinic consultations skyrocketed, filling all 55 designated Kobe hospital bed within 48 hours. Inquiries at call centers increased more rapidly than numbers of subjects rushing to fever clinics. Just after designated hospital beds were filled, medical service restrictions were rapidly relaxed. Our experiences suggest that compulsory hospitalization broke down quickest in the fever case overflow, so medical call centers may be crucial in preventing fever clinic overflows by subjects with fever of unknown origin not recommended to consult fever clinics. Those with severe influenza symptoms should be given priority in hospitalization and flexible policies are recommended.

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A phase II clinical trial designed as a multicenter open study with 30 participating institutions in Japan was conducted in 167 of 181 infants aged 2 to 6 months to evaluate heptavalent pneumococcal conjugate vaccine (7vPnCV) immunogenicity and safety. From September 22, 2004, to September 16, 2006, of 181 infants registered 167 meeting inclusion criteria were include in the immunogenicity analysis. Among the subjects include in the immunogenicity analysis after primary immunization (3 doses), the percentage with IgG antibody levels of at least 0.5μg/mL ―a primary endpoint of this study―was 94.6％ or higher. In the same cohort after the booster immunization (dose 4), the percentage was 96.0％ or higher. Serious adverse event whose causal relationship to 7vPnCV vaccination could not be ruled out were fever of 38℃ and urticaria in 1 each subject, in whom the study was thus discontinued prematurely. Major adverse systemic post-vaccination events were fever, irritability and somnolence. Major local adverse events were redness, swelling or induration, and pain. All adverse events were transient and recovered spontaneously. The vaccineʼs immunogenicity and safety profile are therefore favorable following primary and booster immunization and compatible with those reported in overseas studies. 7vPnCV is expected to show the safety and efficacy similar to that in other countries and to reduce pneumococcal disease in Japan.

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Candida glabrata was continuously isolated in cultured urine samples from a subject with thrombotic thrombocytopenic purpura. Yeast-like fungal phagocytosis found in gram staining led to agents being tested for antifungal susceptibility, revealing hyposensitivity to micafungin (MCFG) of MIC ＜2mg/mL. MCFG administered for 10 days failed to cure C. glabrata infection. To clarify why hyposensitivity occurred, we analyzed the FKS gene sequence using the PCR, finding a deficit of 3 bases coding phenylalanine at FKS2 gene amino acid 659. MCFG hyposensitivity may thus occur in long-term candin-class anti-fungal agent treatment. Candin-class agents have potent anti-fungal activity with fewer adverse effects and are widely used clinically. Hyposensitivity due to resistant C. glabrata species showed thus be considered in fungal infection treatment.

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Kaposiʼs sarocoma (KS) is a well-known complication of the acquired immunodeficiency syndrome (AIDS). A 23-year-old man with AIDS complicated by multiple KS seen in January 2008 for anorexia and 10 kg weight loss had a CD4 cell count of 7/μL and a serum HIV RNA level of 29,000copies/mL. Computed tomography (CT) and endoscopy showed multiple KS lesions in both lungs, the duodenum, small intestine, colon, liver, and both kidneys but not of the skin. Despite the administration of pegylated liposomal doxorubicin (PLD) and highly active antiretroviral therapy, he died in disease progression, unable to complete PLD,KS-related respiratory failure.

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