The purpose of this study is to reevaluate the sensitivities of different methods used in the diagnosis of measles including virus isolation, RT-PCR, and measurement of IgM. Sixty-three throat swabs, 84 peripheral blood mononuclear cell (PBMC) samples, and 85 plasma samples were collected from 85 cases of suspected measles. The sensitivity of virus isolation using throat swabs and PBMC in comparison with RT-PCR was 58.1 and 93.5%, respectively. We defined laboratory-confirmed cases as those in which at least one of the methods was positive. The percentage of positive results by the different methods was compared among 49 laboratory-confirmed cases. The percentage of positive results from PBMC by RT-PCR and virus isolation was 100 and 91.7%, respectively. The percentage of positive results from throat swabs by RT-PCR and virus isolation was 91.2 and 52.8%, respectively. The percentage of IgM positive (79.6%) was significantly lower than that of PBMC by RT-PCR. Ten of 27 plasma samples collected within 5 days of the onset of fever were IgM negative. In contrast, all of the 21 plasma samples collected 6 days after the onset of fever were IgM positive. In conclusion, the detection of measles virus RNA in PBMC by RT-PCR was the most effective method for diagnosis of measles.

Hand-foot-and-mouth disease (HFMD) is caused by a group of enteroviruses, most commonly coxsackievirus A 16 (CA16) and enterovirus 71 (EV71). In general, the disease is mild and self-limited except in the case of EV71 infections, which may incur serious complications. This research focused on virus characterization of HFMD cases in Thailand from 2008–2009, related clinical findings and complications of specific enterovirus subtypes. Specimens (stool, vesicle fluid, throat swab/sputum) from 48 cases were collected during 2008–2009. Reverse transcriptase-polymerase chain reaction (PCR) followed by direct sequencing and phylogenetic analysis served to detect enterovirus and determine subtype. Enterovirus was found in 58.3% (28/48) of cases, specifically EV71 (n=23), CA16 (n=4), and CA10 (n=1). Two patients infected by EV71 had brainstem encephalitis (one death). Eight patients required hospital admission due to dehydration. Of these, 3 were PCR positive for EV71, 1 for CA16, and the reminder negative. This study demonstrated EV71 as the most prevalent present cause of HFMD in Thailand in 2008–2009. Potentially fatal complications of HFMD should be taken into consideration. Surveillance of epidemiology and monitoring of disease severity should be continued, and as a prevention measure, sanitation and hygiene should be improved.

We evaluated the Crystal VC(R), a commercially produced dipstick, for the rapid detection of Vibrio cholerae serotypes O1 and O139 directly from the stool samples of hospitalized diarrheal patients using the conventional bacteriological method as gold standard. The sensitivity and specificity of the dipsticks were about 92 and 73%, respectively. Introduction of the PCR-based method along with the classical bacteriological method as the gold standard for the evaluation of a kit may improve the sensitivity and specificity of the assay. The dipstick method requires minimal technical skill, and the test can be read in about 10 min. This dipstick test has the potential to act as an early warning system for cholera in many developing countries, especially during the start of an outbreak, which would ultimately lead to a decrease in the spread of the disease as well as the case fatality rate. Furthermore, the use of a rapid detection test will improve surveillance and thus reduce the burden of disease estimates, especially in remote settings.

A total of 19,973 clinical specimens obtained from suspected cases of pandemic influenza A virus infection were analyzed by real-time reverse transcription-polymerase chain reaction. Mutations in hemagglutinin (HA) gene and alteration at position 275 in neuraminidase (NA) gene of the randomly selected 29 isolates were detected by sequencing analysis. The virus RNA was detected in 47.3% of the clinical specimens. The pandemic flu cases increased from the 42nd week and peaked in the 46th week of 2009. This intensity continued to the end of the study period. Pandemic flu mainly affected children in the 5–14 year age group, without any gender predominance. The analyzed strains had >98.9% homology with vaccine strains and with each other. More than 37% of the isolates had mutation at position D222E/N on HA gene. There was no isolate harbored mutation at the position H275Y of the NA gene, indicating that the virus isolates currently circulating in Turkey are sensitive to oseltamivir.

A novel method for purifying DNA from clinical samples based on the complex formation of DNA and magnesium ion (Mg²⁺) was developed for the detection of Mycobacterium tuberculosis. The formation of a DNA-Mg²⁺ complex under alkaline conditions was observed by analyzing electrophoretic mobility reduction of DNA on agarose gel. The DNA-Mg²⁺ complex increases the efficacy of DNA recovery from the sample solution on polyethylene terephthalate non-woven fabric (PNWF) filters. Among the various divalent metal cations, only Mg²⁺ was associated with this effect. The applicability of DNA recovered on the PNWF filter was examined for the gene amplification method; loop-mediated isothermal amplification (LAMP). DNA on the PNWF filter could be used for the amplification of specific DNA fragments without elution from the filter. Using this method, DNA was directly purified from M. tuberculosis spiked sputum and examined by LAMP assay, showing a high sensitivity in comparison to the commercially available DNA extraction kit. These results indicated that the method developed in this study is useful for rapid gene diagnosis of tuberculosis.

In July 2009, a pandemic influenza (H1N1) (pdm H1N1) virus epidemic emerged rapidly in Phitsanulok, Thailand. Adult cases of community-acquired pneumonia (CAP) were prospectively examined for pdm H1N1 virus infections by real-time PCR in a tertiary hospital in Phitsanulok from July to November 2009. Twenty-four cases of pdm H1N1 virus-associated CAP were confirmed, and their clinical features including bacterial infection, severity of disease, course of admission, treatment, and outcome were investigated. The median age of these cases was 39.5 years. Most cases appeared to be primary viral pneumonia, but only one case was positive for a urinary pneumococcal antigen. The median time from the onset of illness to admission was 4 days. All 24 patients received oseltamivir after admission. Twelve (50.0%) were defined as having severe CAP and 9 (37.5%) were diagnosed with acute respiratory distress syndrome (ARDS). During the study period, pdm H1N1 virus infections frequently caused severe CAP among young adults because of the delayed initiation of antiviral therapy. Of the 9 ARDS patients, 3 died of ventilator-associated pneumonia caused by multidrug-resistant Acinetobacter baumannii. Implementation of infection control targeting this pathogen is required in tertiary hospitals in Thailand.

Staphylocoagulase (SC) is a major phenotypic determinant of Staphylococcus aureus. Antigenic specificity of SC (SC serotype) has been classified into at least 10 types and employed as an epidemiological marker. In the present study, from the sequence information of SC genes, a novel multiplex PCR assay was developed to determine SC genotypes (SC types) I–X corresponding to SC serotypes I–X, respectively, and to discriminate two subtypes of SC types IV and V. Two PCR reactions (Sc-R1 and Sc-R2) for a single isolate were designed for assigning common SC types in S. aureus isolates from humans (I–VIII and X). When amplicon was not produced, an additional PCR (Sc-R3) was performed to assign SC type IX or subtype Vb. Subtypes IVa and IVb were discriminated by an additional PCR (Sc-R4). SC types were discriminated successfully for the S. aureus strains with established SC types I–X including two subtypes of IV and V. The multiplex PCR assay established in this study could assign SC types for S. aureus clinical isolates at a high determination rate, providing more accurate information on incidence of SC types, and was considered to be useful for epidemiologic characterization of S. aureus.

Bacillary dysentery is still a common and serious public health problem in China. This paper is aimed at developing and evaluating an innovative hybrid model, which combines the seasonal autoregressive integrated moving average (SARIMA) and the generalized regression neural network (GRNN) models, for bacillary dysentery forecasting. Data of monthly bacillary dysentery incidence in Yichang City from 2000–2007 was obtained from Yichang Disease Control and Prevention Center. The SARIMA and SARIMA-GRNN model were developed and validated by dividing the data file into two data sets: data from the past 5 years was used to construct the models, and data from January to June of the 6th year was used to validate them. Simulation and forecasting performance was evaluated and compared between the two models. The hybrid SARIMA-GRNN model was found to outperform the SARIMA model with the lower mean square error, mean absolute error, and mean absolute percentage error in simulation and prediction results. Developing and applying the SARIMA-GRNN hybrid model is an effective decision supportive method for producing reliable forecasts of bacillary dysentery for the study area.

In clinical cholera, a 3-day course of antibiotic complements extensive rehydration therapy by reducing stool volume, shortening the illness, and averting death. However, antibiotic therapy, which has lifesaving implications for cholera, is often hindered due to multidrug resistance in Vibrio cholerae, the cause of cholera. Crude aqueous mixture and water soluble methanol extract from leaf and bark of Psidium guajava, a tropical fruit guava of the family Myrtaceae, showed strong antibacterial activity against multidrug-resistant V. cholerae O1. The in vitro minimum inhibitory concentration of the crude aqueous mixture and water soluble methanol extract, which was bactericidal against 10⁷ CFU/mL of V. cholerae was determined to be 1,250 μg/mL and 850 μg/mL, respectively. The antibacterial activity of P. guajava was stable at 100ºC for 15–20 min, suggesting nonprotein nature of the active component. The growth of V. cholerae in rice oral rehydration saline (ORS) was completely inhibited when 10 mg/mL (wt/vol) of crude aqueous mixture was premixed with the ORS in a ratio of 1:7 (vol. extract/vol. ORS). P. guajava, which is widely distributed in Bangladesh, thus offers great potential for use in indigenous, herbal medicine for controlling epidemics of cholera.

The purpose of our study was to determine the prevalence of macrolide resistance in 3,188 pharyngeal Streptococcus pyogenes isolates collected at the Institute of Public Health of Serbia during the period 2006–2008. The disk diffusion tests were used to determine the susceptibility of the isolates. Two hundred and sixteen S. pyogenes isolates (6.8%) were resistant to erythromycin, with 9 isolates coresistant to tetracycline: 181 isolates harbored mefA gene, 19 ermB gene, 11 ermA(TR) gene, 5 ermB and mefA genes, and 7 tetM gene. Although the prevalence of macrolide resistance in pharyngeal S. pyogenes isolates is low in Serbia, monitoring of the emergence of resistance is advisable.

We investigated the microbial contamination of 0.02% benzalkonium chloride solution used in catheter kits for intermittent self-catheterization. Of 20 samples examined, 12 (60.0%) were contaminated with 8.8x10²-3.1x10⁶ colony-forming units (cfu)/mL. The contaminants were Pseudomonas fluorescens, Burkholderia cepacia, and Aeromonas spp. These results showed that 0.02% benzalkonium chloride solution used for the lubrication/disinfection of catheters for self-catheterization is susceptible to contamination. Therefore, the lubricant/disinfectant for catheters for self-catheterization was changed from 0.02% benzalkonium chloride solution to 84–87% glycerin containing 0.02% benzalkonium chloride, and microbial contamination of the latter in catheter kits for self-catheterization was reinvestigated. Of 42 samples, 5 (11.9%) were contaminated with 20-2.0x10⁴ cfu/mL. However, the rate of contamination of 84–87% glycerin containing 0.02% benzalkonium chloride was significantly lower than that of 0.02% benzalkonium chloride solution (P<0.0001). The contaminant of 84–87% glycerin containing 0.02% benzalkonium chloride was Bacillus spp. in all contaminated samples. In this survey, neither contaminants of 0.02% benzalkonium chloride solution nor the contaminant of 84–87% glycerin containing 0.02% benzalkonium chloride were the causative microbial species of urinary tract infection.

This study aimed to determine the genotypes of NSP4 in children with acute rotavirus diarrhea and evaluate serum antibody titers to NSP4 in different age groups in Shanghai, China. A total of 171 stool specimens were collected from hospitalized patients ≤5 years of age who had acute rotavirus diarrhea between January 2003 and December 2006. Serum samples were collected from healthy individuals, including 200 for 0–60 months of age and 30 for over 5 years of age. NSP4-B type was the single predominant genotype during 2003–2006 in Shanghai. The titers of NSP4 specific IgG antibody increased with age after birth and peaked during 12–23 months of age, thereafter dropping to a level as low as that in the first 5 months of age. However, high levels of antibody against whole rotavirus were maintained in older children over 5 years of age and in adults. Information on prevalence of NSP4 genotypes in this area of China provides useful data for formulating vaccine policy. Short antibody immune memory compared with that induced mainly by rotavirus structural proteins indicated the protective effect against rotavirus may not persist long term if a single NSP4 protein is applied as the rotavirus vaccine.

To examine whether there are any differences in the rates of HIV-spousal transmission between those who have acquired the virus through blood or through sex in central China. A total of 650 HIV-infected individuals were enrolled, 420 of them were either former commercial plasma donors or recipients of blood (blood transmission group [BTG]), and 230 had acquired HIV infection through sex (sex transmission group [STG]). The spousal transmission rate of HIV was 2% per year (94/420, 11.2 years) in BTG and 8.9% per year (115/230, 5.6 years) in STG. There was a significantly higher transmission rate of HIV through male-to-female (11.7% per year, 84/128, 5.6 years) than through female-to-male (5.4% per year, 31/102, 5.6 years, P<0.05) in STG, but there was no significant gender based differences in BTG. In BTG, all HIV-1 tested were subtype B', while in STG, HIV-1 tested were predominantly subtypes CRF01_AE or CRF07_BC. Our results show that the HIV-spousal transmission rate was higher in STG than in BTG, and that there was a higher rate through male-to-female than female-to-male in STG.

Mupirocin is used topically to treat skin infection caused by methicillin-resistant Staphylococcus aureus (MRSA). One hundred eighty-eight strains (isolated in 2003, 2004, 2007, and 2008) were tested for mupirocin susceptibility using disk diffusion method and minimum inhibitory concentration (MIC). Mupirocin resistance was detected in 10 (5%) strains with 2 of them showing MIC of 256 mg/l. PCR detection using gene-specific primers showed that all 10 mupirocin-resistant strains harbored ileS2 gene whereas mupA gene was detected in 2 mupirocin-resistant strains with MIC of 256 mg/l. Amplification of agr grouping and SCCmec typing showed that all 10 strains were agr group I and SCCmec type III. Sequence analysis of region X of the spa gene yielded 4 distinct spa types (t037, t363, t421, and t6405) which were clonally related. In conclusion, the rate of mupirocin resistance in Malaysia is still low but is much higher than previous reports in Malaysia.

Pantoea agglomerans is a plant pathogen and an unusual cause of human disease typically associated with thorn prick injuries or contaminated parenteral fluids. In the neonate, P. agglomerans has been reported to cause bacteremia or sepsis 17 times previously. In most of these cases, the source of infection has been contaminated parenteral nutrition or has been associated with indwelling catheters. We present a rare case of P. agglomerans bacteremia in a 35-week-female born via vaginal delivery complicated by prolonged rupture of membranes and chorioamnionitis. English language literature on the subject is also reviewed.

Salmonella enterica serovar Typhimurium strains of particular phage types, such as DT104, U302, DT120, DT193, and nontypable strains, are often characterized by resistance to multiple antibiotics. This antibiotic resistance can be caused by the presence of the integrons, transposons, Salmonella genomic island 1 (SGI1), or conjugative plasmids. In this study we were interested in the relative contribution of integrons and SGI1 to the antibiotic resistance of the four mentioned phage types and nontypable S. Typhimurium human strains. Altogether 193 isolates were characterized for antibiotic susceptibility, presence of class 1 integrons, and the left junction of SGI1. Based on the presence of class 1 integrons and the left junction of SGI1, all strains could be clustered into three groups. The first group consisted of 69 strains positive for both the class 1 integrons and the left junction of SGI1. The strains of this group belonged mainly to DT104, U302, and DT120 phage types with resistance phenotype ACSSuT or ACSSuTNA. The second group comprised 9 strains which were positive only for the presence of class 1 integrons. In this group were some strains of multiple-antibiotic-resistant phage types: DT120, DT193, U302; and nontypable. The third group consisted of 115 strains in which neither the class 1 integrons nor the left junction of SGI1 were detected. Although the isolates were resistant to 2–8 antibiotics, the most frequent resistance type of these strains were ASSuT and SSu. By nucleotide sequencing of class 1 integrons PCR amplicons, the following embedded gene cassettes were determined: aadA1, aadA2, aadA5, aadA7, bla_PSE-1, sat1, dfrA1; and dfrA14. Our study shows a high prevalence and diversity of class 1 integrons embedded antimicrobial gene cassettes and their strong association with SGI1.

An ELISA system for measuring antibodies to nonstructural protein 1 (NS1) of Japanese encephalitis virus has already been established. This system uses an ELISA diluent containing casein, instead of bovine serum albumin (BSA). During a survey, we found that 21 (21%) of 102 children aged 1–5 years, who had no history of Japanese encephalitis vaccination and were without detectable neutralizing antibodies, showed positive results with this ELISA system. Western blotting analysis showed that sera from 19 (91%) of these 21 subjects had antibodies to BSA, but not NS1. These sera reacted with BSA antigen remaining in immunoaffinity-purified NS1 antigen. One solution to this problem was to reduce the BSA level to ≤1% of the NS1 amount. Another was to use a control well sensitized with BSA with the same amount as that contained in the NS1 antigen preparation.