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The most frequent markers of fluoroquinolone resistance in S. pneumoniae are chromosomal mutations in the quinolone-resistance-determining regions of DNA gyrase and topoisomerase IV encoding for the gyrA, gyrB and parC, parE genes. In 2008, 6.5% of the Streptococcus pneumoniae isolates in a Bangkok university hospital were resistant to ofloxacin. Using PCR and DNA sequencing, we identified mutations in both the gyrA and parC genes of four ofloxacin- and ciprofloxacin-resistant S. pneumoniae isolates (minimum inhibitory concentrations > 32 microg/ml). Mutations were found in the gyrA gene at positions Ser81Phe, Glu85Gly, Glu85Lys and in the parC gene at position Ser79Tyr. Three isolates had mutations in both genes. Two of the isolates were serotype 6B and two were serotypes not contained in currently licensed pneumococcal vaccines. This is the first report of the mechanisms of fluoroquinolone resistance in S. pneumoniae in Thailand.

Streptococcus pneumoniae was isolated from 170 patient specimens at Siriraj Hospital during January-December 2008. Patients were 66% male and ranged in age from 3 months to 94 years (mean +/- SD = 38.2 +/- 31.7). The largest proportion (29.4%) of isolates were from patients older than 60 years, followed by patients aged 2-5 years (20%) and from patients less than 2 years (12.4%). Monthly isolation was highest in December (22 isolates in December compared to the average of 13 isolates of the other months). Antimicrobial susceptibility for eight drugs was determined by the disk diffusion method. Overall, susceptibility was generally high to chloramphenicol (71.8%), linezolid (100%), ofloxacin (93.5%) and vancomycin (100%), but less susceptible to erythromycin (35.3%), penicillin (31.1%), tetracycline (28.8%) and trimethoprim/ sulfamethoxazole (24.1%). Among the 105 (62%) isolates resistant to three or more drugs, the most common resistance pattern was erythromycin-penicillin-tetracycline-trimethoprim/sulfamethoxazole, accounting for 39% of such isolates, followed by chloramphenicol-erythromycin-penicillin-tetracycline- trimethoprim/sulfamethoxazole (29.5%). The minimal inhibitory concentrations (MIC) of penicillin and cefotaxime were determined by broth microdilution. By 2008 CLSI criteria, 92% and 90% of 51 sterile site isolates were penicillin and cefotaxime susceptible, including one of two meningitis cases. In contrast, of 26 non-sterile site isolates, only 26.9% and 76.9% were susceptible to penicillin and cefotaxime, respectively. The MICs of penicillin were higher for isolates from non-sterile sites than for those from sterile sites.

Rapid detection of bacterial pathogen causing meningitis is very important to guide antimicrobial therapy before the standard culture result is available. Other than gram stain, one of the most useful rapid methods is the detection of bacterial antigen in cerebrospinal fluid. This article reviewed the methods of bacterial antigen detection for diagnosis of meningitis as well as a microbiology aspect of this life-threatening disease.

Objective: Penicillin resistance in Streptococcus pneumoniae isolates results from altered penicillin-binding proteins (PBPs), especially PBP2, which has a reduced affinity to penicillin. This study evaluated drug resistance and the gene sequence of the conserved motif pbp2b of penicillin-resistant isolates in Thailand., Material and Method: Penicillin-resistant pneumococci with minimum inhibitory concentrations (MIC) for penicillin > or = microg/ ml and penicillin-susceptible strains were identified from clinical specimens. The pbp2b genes were amplified by polymerase chain reaction (PCR), and the purified PCR product was cloned into E. coli. The recombinant plasmid clones containing pbp2b were sequenced and evaluated for mutations corresponding to penicillin and cefotaxime resistance., Results: Penicillin-susceptible S. pneumoniae isolates were susceptible to 12 other antibiotics tested (range 95-100%) while penicillin-nonsusceptible isolates were resistant to most antibiotics except amoxicillin/clavulanate and levofloxacin. Sequence analysis of pbp2b showed a substitution of A for T451 next to the region of the SSN triad in all six resistant isolates tested and mutations clustered around the KTG triad in two isolates. Using the ClustalW alignment program, Thai isolates differed from those of European countries, but were more similar to those from Japan than Korea., Conclusion: Penicillin or cefotaxime resistance in S. pneumoniae in Thailand was due to affinity reduction of PBP2b, similar to changes found in other Asian isolates.

Objective: Extended-spectrum beta-lactamases (ESBLs) are most prevalent in Klebsiella pneumoniae. This organism is frequently isolated from clinical specimens and can cause septicemia, pneumonia or urinary tract infection. There were occasionally suspicious outbreaks of ESBL-producing K. pneumoniae in patients' wards. The objective is to determine whether the randomly amplified polymorphic DNA (RAPD), which is a polymerase chain reaction (PCR)-based typing technique, can be used as a typing method for studying the molecular epidemiology of ESBL-producing K. pneumoniae., Material and Method: The present study was carried out by using 30 ESBL-producing K. pneumoniae isolates obtained from different patients admitted to Siriraj Hospital between January and February 2004. RAPD was evaluated for three primers. All isolates were re-examined by using Southern blot hybridization., Results: It was found that 29 DNA band patterns were generated individually by either AP4 or HLWL74 and R108 primers (30 patterns) for RAPD analysis and 30 patterns for Southern blot hybridization with class 1 integron (intI1) probe. Different patterns indicated that these 30 isolates could not be the cause of an outbreak in Siriraj Hospital., Conclusion: The RAPD typing is good and can be used as a screening, rapid and inexpensive'test for ESBL-producing K. pneumoniae during investigation of outbreaks.

Background: The CA-MRSA infections have emerged in many parts of the world over the past decade. To our knowledge, the prevalence of CA-MRSA infections in Thai patients is unknown., Objective: To determine an epidemiology of Staphylococcus aureus (S. aureus) infections in hospitalized patients in Siriraj Hospital and the prevalence of infections caused by community-acquired methicillin-resistant S. aureus (CA-MRSA)., Material and Method: The study was carried out at Siriraj Hospital from January to May 2005. The eligible patients were hospitalized patients whom S. aureus were isolated from their clinical specimens submitted to Department of Microbiology. S. aureus isolate was classified into infection or colonization. S. aureus infections were further classified into methicillin-resistant S. aureus (MRSA) or methicillin-sensitive S. aureus (MSSA) infections, and hospital-acquired (HA) or community-acquired (CA) infections. CA-MRSA infection is defined as infection caused by MRSA isolated from the patient within 72-hour of hospitalization and has no features of HA MRSA infections., Results: There were 669 S. aureus isolates from 448 patients. Two hundred and sixty two patients (58.5%) were MSSA whereas 186 (41.5%) were MRSA infections. CA-MRSA was found in three isolates (0.9% of total MRSA) from two patients., Conclusion: The prevalence of CA-MRSA infections in hospitalized patients in Siriraj Hospital was uncommon and these patients could probably be HA MRSA infections.

Mycoplasma pneumoniae antibody was determined in 811 sera of different patients admitted to Siriraj Hospital with respiratory tract infection from July 1, 2000 to August 31, 2003 by agglutination with gelatin particle agglutination test kit (SERODIA-MYCO II, Fujirebio Inc. Japan) in microtiter plates. Three hundred and three sera were positive (37.36%). The five most positive titer were found in patients 5-9 yr (40.26%), followed by patients 1-4 yr (24.75%), 10-14 yr (19.80%), 30-39 yr (5.28%) and 20-29 yr (3.96%). The positive titers ranged from 40 to > 20,480. Female:male ratio in positive patients was approximately the same (1.19:1). High titers (> or = 320) were found in 146 out of 303 patients (48.18%). The infection was mostly found in children aged 5-9 yr. Detection of antibody to M. pneumoniae infection showed that 37.36% of patients who were suspected of having atypical bacterial pneumonia were positive.

A prospective study in pediatric patients compared the BACTEC system, an automated blood culture system using Bactec Ped Plus/F broth media, with the conventional system using Brain Heart Infusion broth media. Each single blood sample drawn for culture from hospitalized children was evaluated by both systems simultaneously. Of 244 pairs of blood samples, 44 (18%) were positive for microorganisms. Of these isolates, 24 (55%) were detected by both systems, 10 (23%) were detected by the BACTEC system only, and 10 (23%) were detected by the conventional system only. The mean turn around time of the BACTEC system (0.56 +/- 0.34, range 0.08-1.55 days) was significantly shorter than that of the conventional system (3.36 +/- 2.72, range 1-7 days, p < 0.001). Seven isolates strongly suspected to be due to contaminants grew out after 5 days of incubation and were detected by the conventional system only. In this study the BACTEC system and the conventional system were equally effective in detecting microorganisms in the patients' blood samples even if antibiotic therapy had been previously administered. However, the results from the BACTEC system were available much sooner and less likely to be contaminants.

Three cases with S. suis bacteremia and meningitis were reported. The first case was a 23-year-old butcher who was a regular drinker of alcohol for two years and developed streptococcal toxic-shock syndrome. The organism was transmitted to him through a minor cut in his right arm. The second cases was a 49-year-old female laborer who had been consuming locally produced alcohol for 20 years and developed fever and meningitis. Unfortunately, she succumbed in seven days despite intensive supportive and cefotaxime treatments. The third case was a 45-year-old regular alcoholic drinker and car painter who was seen at a private hospital due to contusion at his left lateral chest wall. However, fever and confusion due to meningitis was detected upon admission. Irreversible deafness developed within 48 hours of ceftriaxone therapy for meningitis. He finally recovered with deafness. S. suis was isolated from blood and cerebrospinal fluid cultures in all three cases though initially reported to be viridans group of streptococci.