Spread of carbapenem-resistant Acinetobacter baumannii global clone 2 in Asia and AbaR-type resistance islands.

In this surveillance study, we identified the genotypes, carbapenem resistance determinants, and structural variations of AbaR-type resistance islands among carbapenem-resistant Acinetobacter baumannii (CRAB) isolates from nine Asian locales. Clonal complex 92 (CC92), corresponding to global clone 2 (GC2), was the most prevalent in most Asian locales (83/108 isolates; 76.9%). CC108, or GC1, was a predominant clone in India. OXA-23 oxacillinase was detected in CRAB isolates from most Asian locales except Taiwan. blaOXA-24 was found in CRAB isolates from Taiwan. AbaR4-type resistance islands, which were divided into six subtypes, were identified in most CRAB isolates investigated. Five isolates from India, Malaysia, Singapore, and Hong Kong contained AbaR3-type resistance islands. Of these, three isolates harbored both AbaR3- and AbaR4-type resistance islands simultaneously. In this study, GC2 was revealed as a prevalent clone in most Asian locales, with the AbaR4-type resistance island predominant, with diverse variants. The significance of this study lies in identifying the spread of global clones of carbapenem-resistant A. baumannii in Asia.

Dissemination of metallo-ß-lactamase-producing Pseudomonas aeruginosa of sequence type 235 in Asian countries.

OBJECTIVES: To investigate the epidemiological traits of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa (MPPA) clinical isolates collected by the Asian Network for Surveillance of Resistant Pathogens (ANSORP). METHODS: A total of 16 MPPA clinical isolates were collected from six Asian countries in 2000 to 2009 by ANSORP. The MBL gene was detected by PCR amplification. The genetic organization of the class 1 integron carrying the MBL gene cassette was investigated by PCR mapping and sequencing. Southern blotting, repetitive sequence-based PCR and multilocus sequence typing (MLST) experiments were performed to characterize the isolates. RESULTS: PCR and sequencing experiments detected the blaVIM-2 (n = 12), blaVIM-3 (n = 1), blaIMP-6 (n = 2) and blaIMP-26 (n = 1) genes. The MBL genes were located on the chromosome in all isolates except one. Furthermore, all the MBL genes were located in a class 1 integron. All the MPPA isolates from Malaysia, Thailand, Sri Lanka and Korea were identified as sequence type (ST) 235 by MLST. Three VIM-2-producing isolates from India were identified as ST773, and one isolate harbouring VIM-3 from Taiwan was identified as ST298. CONCLUSIONS: P. aeruginosa ST235 might play a role in dissemination of MBL genes in Asian countries.

Changing trends in antimicrobial resistance and serotypes of Streptococcus pneumoniae isolates in Asian countries: an Asian Network for Surveillance of Resistant Pathogens (ANSORP) study.

Antimicrobial resistance in Streptococcus pneumoniae remains a serious concern worldwide, particularly in Asian countries, despite the introduction of heptavalent pneumococcal conjugate vaccine (PCV7). The Asian Network for Surveillance of Resistant Pathogens (ANSORP) performed a prospective surveillance study of 2,184 S. pneumoniae isolates collected from patients with pneumococcal infections from 60 hospitals in 11 Asian countries from 2008 to 2009. Among nonmeningeal isolates, the prevalence rate of penicillin-nonsusceptible pneumococci (MIC, ≥ 4 µg/ml) was 4.6% and penicillin resistance (MIC, ≥ 8 µg/ml) was extremely rare (0.7%). Resistance to erythromycin was very prevalent in the region (72.7%); the highest rates were in China (96.4%), Taiwan (84.9%), and Vietnam (80.7%). Multidrug resistance (MDR) was observed in 59.3% of isolates from Asian countries. Major serotypes were 19F (23.5%), 23F (10.0%), 19A (8.2%), 14 (7.3%), and 6B (7.3%). Overall, 52.5% of isolates showed PCV7 serotypes, ranging from 16.1% in Philippines to 75.1% in Vietnam. Serotypes 19A (8.2%), 3 (6.2%), and 6A (4.2%) were the most prominent non-PCV7 serotypes in the Asian region. Among isolates with serotype 19A, 86.0% and 79.8% showed erythromycin resistance and MDR, respectively. The most remarkable findings about the epidemiology of S. pneumoniae in Asian countries after the introduction of PCV7 were the high prevalence of macrolide resistance and MDR and distinctive increases in serotype 19A.

High prevalence of multidrug-resistant nonfermenters in hospital-acquired pneumonia in Asia.

RATIONALE: Hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP) remain important causes of morbidity and mortality. Increasing antimicrobial resistance has aroused the concern of the failure of antibiotic treatment. OBJECTIVES: To determine the distribution of the bacterial isolates of HAP and VAP, their antimicrobial resistance patterns, and impact of discordant antibiotic therapy on clinical outcome in Asian countries METHODS: A prospective surveillance study was conducted in 73 hospitals in 10 Asian countries from 2008-2009. A total of 2,554 cases with HAP or VAP in adults were enrolled and 2,445 bacterial isolates were collected from 1,897 cases. Clinical characteristics and antimicrobial resistance profiles were analyzed. MEASUREMENT AND MAIN RESULTS: Major bacterial isolates from HAP and VAP cases in Asian countries were Acinetobacter spp., Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae. Imipenem resistance rates of Acinetobacter and P. aeruginosa were 67.3% and 27.2%, respectively. Multidrug-resistant rates were 82% and 42.8%, and extensively drug-resistant rates were 51.1% and 4.9%. Multidrug-resistant rate of K. pneumoniae was 44.7%. Oxacillin resistance rate of S. aureus was 82.1%. All-cause mortality rate was 38.9%. Discordant initial empirical antimicrobial therapy increased the likelihood of pneumonia-related mortality (odds ratio, 1.542; 95% confidence interval, 1.127-2.110). CONCLUSIONS: Acinetobacter spp., P. aeruginosa, S. aureus, and K. pneumoniae are the most frequent isolates from adults with HAP or VAP in Asian countries. These isolates are highly resistant to major antimicrobial agents, which could limit the therapeutic options in the clinical practice. Discordant initial empirical antimicrobial therapy significantly increases the likelihood of pneumonia-related mortality.

Spread of methicillin-resistant Staphylococcus aureus between the community and the hospitals in Asian countries: an ANSORP study.

OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA) is highly prevalent in hospitals in many Asian countries. Recent emergence of community-associated (CA) MRSA worldwide has added another serious concern to the epidemiology of S. aureus infections. To understand the changing epidemiology of S. aureus infections in Asian countries, we performed a prospective, multinational surveillance study with molecular typing analysis. METHODS: We evaluated the prevalence of methicillin resistance in S. aureus isolates in CA and healthcare-associated (HA) infections, and performed molecular characterization and antimicrobial susceptibility tests of MRSA isolates. RESULTS: MRSA accounted for 25.5% of CA S. aureus infections and 67.4% of HA infections. Predominant clones of CA-MRSA isolates were ST59-MRSA-SCCmec type IV-spa type t437, ST30-MRSA-SCCmec type IV-spa type t019 and ST72-MRSA-SCCmec type IV-spa type t324. Previously established nosocomial MRSA strains including sequence type (ST) 239 and ST5 clones were found among CA-MRSA isolates from patients without any risk factors for HA-MRSA infection. CA-MRSA clones such as ST59, ST30 and ST72 were also isolated from patients with HA infections. CONCLUSIONS: Our findings confirmed that MRSA infections in the community have been increasing in Asian countries. Data also suggest that various MRSA clones have spread between the community and hospitals as well as between countries.

Antimicrobial susceptibility pattern and serotyping of Streptococcus pneumoniae isolated from Kanti Children Hospital in Nepal.

BACKGROUND: Invasive pneumococcal disease is a significant cause of morbidity and mortality worldwide and it is a major cause for childhood deaths in Nepal. OBJECTIVES: The aim of this study was to establish the antimicrobial susceptibility pattern of Streptococcus pneumoniae and perform serotype responsible for pneumococcal disease in Nepal. MATERIALS AND METHODS: All together 3774 children from 2 to 60 months who fulfilled the enrollment criteria for suspect of bacterial pneumonia, sepsis or meningitis were enrolled for etiologic studies of severe illness. During the study period 60 isolates of Streptococcus pneumoniae were isolated and the antimicrobial susceptibility testing and serotyping were performed. RESULTS: The study showed that 24 (52. 17%) isolates were resistant to Cotrimoxazole, 3 (6. 5%) isolates were intermediately resistant to Penicillin but no Penicillin resistant strains were isolated. The 1 (2. 17%) isolate was recorded as Erythromycin and Chloramphenicol resistant and only 1 (2. 17%) isolate was found intermediately resistant to Cefotaxime. Of the 60 isolates, serotyping result was available only for 46 isolates. The most common serotypes were serotype 1 (27. 65%) followed by serotype 5 (19. 14%) and serotype 4 (8. 5%) respectively followed by serotype 39, 23F, 7F, 19B, 12A, 14, 18F, 6B, 32, 16, 19F and 25F. CONCLUSIONS: Alarming level of Cotrimoxazole resistance demands revision of pneumonia treatment policy in Nepal and rising tendency of other drug resistance against Streptococcus pneumoniae showed use of these drugs for the treatment of meningitis, pneumonia and other serious infections needs extended research. The common serotype 1, 5 and 4 need to be incorporated in pneumococcal vaccine to immunise children in Nepal.

Invasive pneumococcal disease in Kanti Children's Hospital, Nepal, as observed by the South Asian Pneumococcal Alliance network.

BACKGROUND: Pneumonia accounts for approximately 2 million deaths annually among children aged <5 years, with most of these deaths occurring in Africa and southern Asia. The South Asian Pneumococcal Alliance (SAPNA) network in Nepal is generating local epidemiological data to assist in the development of national and regional policies for prevention of pneumococcal and Haemophilus influenzae (Hib) disease. METHODS: Children aged 2 months to 5 years with suspected invasive bacterial disease were recruited from Kanti Children Hospital, Kathmandu, Nepal. Specimens of blood, CSF, and normally sterile body fluids were cultured, and analysis of antimicrobial susceptibility patterns and serotyping of Streptococcus pneumoniae isolates were performed. CSF specimens were also tested for S. pneumoniae and Hib antigens by a latex agglutination test and an immunochromatographic test of pneumococcal antigen (NOW S. pneumoniae Antigen Test; Binax). RESULTS: A total of 2528 children with suspected invasive bacterial disease were recruited, of whom 82% had pneumonia, 9.6% had meningitis, 2% had very severe disease, and 0.4% had bacteremia; the remainder received another diagnosis. Before hospitalization, 26.7% had received antibiotic treatment. Fifty children had S. pneumoniae identified as the etiological agent of invasive disease. Of 2461 blood cultures performed, 22 were positive for S. pneumoniae. Of 33 cases of S. pneumoniae meningitis, 11 were detected by CSF culture, and 21 were detected by latex agglutination and pneumococcal antigen tests. The rate of detection of S. pneumoniae in CSF was 3.6% by culture, compared with 7.8% by latex agglutination and 10% by pneumococcal antigen testing. The rate of detection of H. influenzae in CSF was 1.7% by culture and 6.5% by latex agglutination. The most common serotypes found were 1, 5, 2, and 7F, followed by 12A, 19B, and 23F. Of all the invasive isolates, 3.8% were resistant to penicillin, and 68% were resistant to trimethoprim-sulfamethoxazole. CONCLUSIONS: The SAPNA network has identified Hib and pneumococci as causes of significant disease in Nepal.

Surveillance of invasive pneumococcal disease in Colombo, Sri Lanka.

The South Asian Pneumococcal Surveillance network uses standard recruitment and laboratory procedures for surveillance of invasive pneumococcal disease in India, Nepal, and Sri Lanka. Children aged 2 months to 5 years who were admitted to the sentinel surveillance site, Lady Ridgeway Hospital for Children, in Colombo, Sri Lanka, and who presented with signs and symptoms of meningitis, pneumonia, or very severe disease were studied. Blood culture and CSF culture specimens were analyzed at the microbiology laboratory at Lady Ridgeway Hospital for Children. Specimens were processed by routine conventional methods. Antigen testing was performed on CSF specimens with use of commercially available latex agglutination test kits. From January 2005 to March 2007, we observed 23 isolates of Streptococcus pneumoniae, and the most common serotypes were 19F, 14, 23F, and 6B. Of the serotypes found, 60% are covered by the currently available 7-valent conjugate pneumococcal vaccine. More than 90% of the isolates were penicillin resistant, and the rate of resistance to third-generation cephalosporins was also high.

Community effect of Haemophilus influenzae type b vaccination in India.

We assessed the effect of distribution of Haemophilus influenzae type b (Hib) vaccine in the private health care sector on Hib meningitis admissions at a referral hospital in India. The annual mean number of Hib cases was 10.7 before Hib vaccine introduction, falling to 3.8 cases following introduction (P < 0.0001). By contrast, the mean of annual numbers of pneumococcal cases were 3.0 and 4.6, (P = 0.55). Even at relatively low coverage through private sector distribution, Hib vaccine has significant community impact on Hib disease.

Epidemiology and clinical outcomes of community-acquired pneumonia in adult patients in Asian countries: a prospective study by the Asian network for surveillance of resistant pathogens.

Appropriate antimicrobial treatment of community-acquired pneumonia (CAP) should be based on the distribution of aetiological pathogens, antimicrobial resistance of major pathogens, clinical characteristics and outcomes. We performed a prospective observational study of 955 cases of adult CAP in 14 hospitals in eight Asian countries. Microbiological evaluation to determine etiological pathogens as well as clinical evaluation was performed. Bronchopulmonary disease (29.9%) was the most frequent underlying disease, followed by cardiovascular diseases (19.9%), malignancy (11.7%) and neurological disorder (8.2%). Streptococcus pneumoniae (29.2%) was the most common isolate, followed by Klebsiella pneumoniae (15.4%) and Haemophilus influenzae (15.1%). Serological tests were positive for Mycoplasma pneumoniae (11.0%) and Chlamydia pneumoniae (13.4%). Only 1.1% was positive for Legionella pneumophila by urinary antigen test. Of the pneumococcal isolates, 56.1% were resistant to erythromycin and 52.6% were not susceptible to penicillin. Seventeen percent of CAP had mixed infection, especially S. pneumoniae with C. pneumoniae. The overall mortality rate was 7.3%, and nursing home residence, mechanical ventilation, malignancy, cardiovascular diseases, respiratory rate>30/min and hyponatraemia were significant independent risk factors for mortality by multivariate analysis (P<0.05). The current data provide relevant information about pathogen distribution and antimicrobial resistance of major pathogens of CAP as well as clinical outcomes of illness in Asian countries.

Comparison of phenotypic versus genotypic methods in the detection of methicillin resistance in Staphylococcus aureus.

BACKGROUND & OBJECTIVE: Conventional methods to detect methicillin resistance in Staphylococcus aureus are inadequate as expression of resistance is subject to environmental and conditional expression of PBP2a antigen. The objective of the present study was to determine methicillin resistance in S. aureus by conventional susceptibility (oxacillin disc diffusion and oxacillin MIC) and molecular methods (PCR) and to evaluate latex agglutination test for the detection of PBP 2a and to compare the results of these tests for its sensitivity, specificity and rapidity. METHODS: A total of 150 consecutive clinical isolates of Staphylococcus aureus received at the Department of Microbiology, Christian Medical College, Vellore, were included. Oxacillin (1 mg) disc diffusion and agar dilution method were used. The isolates were also subjected to latex agglutination test for detection of PBP2a and multiplex PCR to detect mecA and femB genes. RESULTS: Of the 150 isolates, 33 were found to be MRSA by oxacillin disc diffusion. By MIC method, 13 per cent of the isolates had values 32 microg/ml, 6 per cent between 16-8 mug/ml and 2.7 per cent had a value of 4 microg/ml; 100 per cent concordance was obtained between the oxacillin disc screening and MIC methods. The latex agglutination showed positive reaction for all MRSA with only one MSSA being falsely classified as MRSA. The specificity and sensitivity were 99 and 100 per cent respectively. Test results were obtained within 15 min. By multiplex PCR, all 22 per cent of MRSA were positive for mecA and femB genes and additionally one MSSA carried mecA gene. However, femB gene was not found in 6 MSSA isolates. Specificity and sensitivity of PCR for mecA detection was similar to latex agglutination test. PCR system required approximately five hours. INTERPRETATION & CONCLUSION: Our findings showed that the conventional methods for detection of methicillin resistance like disc screening, disc diffusion and MIC are cost-effective but time consuming. Latex agglutination though expensive is rapid and can be a good preliminary screen with high sensitivity and specificity. Multiplex PCR is a good confirmatory test though expensive.

Incidence of Haemophilus influenzae type b meningitis in India.

BACKGROUND & OBJECTIVE: Vaccine policy depends on locally relevant disease burden estimates. The incidence of Haemophilus influenzae type b (Hib) disease is not well characterized in the South Asian region, home to 30 per cent of the world's children. There are limited data from prospective population incidence studies of Hib in Asia, and no data available from India. We therefore carried out this study to assess the burden of Hib meningitis in India. METHODS: A prospective surveillance study was carried out during 1997 and 1999 in hospitals for cases of Hib meningitis from 5 administrative areas of an Indian district (Vellore, Tamil Nadu) with 56,153 children under 5 yr of age, over a 24 month period RESULTS: Ninety seven cases of possible meningitis (> 10 WBC/microl in CSF) were reported, an annual incidence of 86 per 100,000 (95%CI 69 to 109) in 0-4 yr old children, and 357 per 100,000 in 0-11 month infants. Eighteen had proven bacterial meningitis, an annual incidence of 15.9 per 100,000. Eight CSF had Hib by culture or antigen testing, an annual incidence of 7.1 per 100,000 (95%CI 3.1 to 14.0) in children 0-59 months. In infants 0-11 months of age, the incidence of Hib meningitis was 32 per 100,000 (95%CI 16 to 67) and in the 0-23 month group it was 19 (95%CI 8 to 37). INTERPRETATION & CONCLUSION: Our data are the first minimal estimate of the incidence of Hib meningitis for Indian children. The observed incidence data are similar to European reports before Hib vaccine use, suggest substantial disease before 24 months of age, and provide data useful for policy regarding Hib immunization.

Extended-Spectrum Beta-lactamase Producers: Detection for the Diagnostic Laboratory.

BACKGROUND AND OBJECTIVES: Discovered in 1983, Extended spectrum beta-lactamase (ESBL) producers are still the leading cause of infections in India. Its prompt detection is crucial to the clinical management. The Clinical Laboratory Standards Institute (CLSI) recommends phenotypic screening and confirmatory tests to identify the ESBL producer making it cost and time consuming for the diagnostic laboratory. We compare here the screening and confirmatory tests offering a solution to the CLSI recommendation. METHODS: Nosocomial isolates E. coli (71) and K. pneumoniae (25) resistant to cefotaxime and ceftazidime were included. CLSI recommended testing with cefotaxime, ceftazidime and in combination with clavulanic acid by disk diffusion and agar dilution methods were performed. E-test was performed on discrepant results. To determine the genetic relatedness of the organisms, 22 Medical and Surgical ICU isolates were genotyped by PFGE. Dendrogram was constructed using dice co-efficient, UPGMA method with diversity database software. RESULTS AND CONCLUSIONS: Phenotypic screening disk diffusion test versus the confirmatory agar dilution MIC tests with cefotaxime and ceftazidime correlated well with the final ESBL status (kappa 0.852 and 0.905 P < 0.001) and (kappa 0.911 and 0.822 P < 0.001). The tests show 99-100% sensitivity, 75-83.3% specificity, and positive likelihood ratios between 4.0 -5.9. E-test confirmed 6 of 12 discordant results as ESBLs. Of the 96 nosocomial isolates screened as possible ESBL producers by the Kirby-Bauer disk diffusion test, 86.5% were confirmed ESBL producers. Genotyping on the ICU isolates by PFGE revealed a genetically diverse population suggesting no transmission of phenotypically similar ESBL strains within the ICUs.

Secondary sternal Aspergillus osteomyelitis in a diabetic hemodialysis patient with previous allograft rejection.

A 29-year-old diabetic woman who had a previously failed renal allograft on maintenance hemodialysis developed sternal aspergillosis with Aspergillus terreus following a pericardiectomy. She was successfully treated with surgical debridement and a combination of antifungal agents including amphotericin B, caspofungin, and voriconizole. The diagnostic difficulties and management are discussed.

Reproductive tract infections among young married women in Tamil Nadu, India.

CONTEXT: Women often suffer silently with reproductive tract infections (RTIs). Studies of the prevalence of these infections in South Asia have been hindered by low participation rates, and little is known about rates among the youngest married women. METHODS: A community-based cross-sectional study of RTIs was conducted in 1996-1997 among married women 16-22 years of age in Tamil Nadu, India. The women were questioned about symptoms, received pelvic and speculum examinations and provided samples for laboratory tests. Qualitative and quantitative data on treatment-seeking behavior were collected. RESULTS: Fifty-three percent of women reported gynecologic symptoms, 38% had laboratory findings of RTIs and 14% had clinically diagnosed pelvic inflammatory disease or cervicitis. According to laboratory diagnoses, 15% had sexually transmitted infections and 28% had endogenous infections. Multivariate analysis found that women who worked as agricultural laborers had an elevated likelihood of having a sexually transmitted infection (odds ratio, 2.4), as did those married five or more years (2.1). Two-thirds of symptomatic women had not sought any treatment; the reasons cited were absence of a female provider in the nearby health care center, lack of privacy, distance from home, cost and a perception that their symptoms were normal. CONCLUSIONS: Young married women in this rural Indian community have a high prevalence of RTIs but seldom seek treatment. Education and outreach are needed to reduce the stigma, embarrassment and lack of knowledge related to RTIs. The low social status of women, especially young women, appears to be a significant influence on their low rates of treatment for these conditions.

An outbreak of cutaneous anthrax in a non-endemic district--Visakhapatnam in Andhra Pradesh.

BACKGROUND: Anthrax is a disease of herbivorous animals, and humans incidentally acquire the disease by handling infected dead animals and their products. Sporadic cases of human anthrax have been reported from Southern India. METHODS: Five tribal men presented with painless ulcers with vesiculation and edema of the surrounding skin on the extremities without any constitutional symptoms. There was a history of slaughtering and consumption of a dead goat ten days prior to the development of skin lesions. Clinically cutaneous anthrax was suspected and smears, swabs and punch biopsies were taken for culture and identification by polymerase chain reaction (PCR). All the cases were treated with intravenous followed by oral antibiotics. Appropriate health authorities were alerted and proper control measures were employed. RESULTS: Smears from the cutaneous lesions of all five patients were positive for Bacillus anthracis and this was confirmed by a positive culture and PCR of the smears in four of the five cases. All the cases responded to antibiotics. CONCLUSION: We report five cases of cutaneous anthrax in a non-endemic district, Visakhapatnam, Andhra Pradesh, for the first time.

Clinical outcomes of pneumococcal pneumonia caused by antibiotic-resistant strains in asian countries: a study by the Asian Network for Surveillance of Resistant Pathogens.

To evaluate the clinical outcomes of pneumococcal pneumonia caused by antibiotic-resistant strains in Asian countries, we performed a prospective observational study of 233 cases of adult pneumococcal pneumonia in 9 Asian countries from January 2000 to June 2001. Among 233 isolates, 128 (55%) were not susceptible to penicillin (25.3% were intermediately susceptible, and 29.6% were resistant). Clinical severity of pneumococcal pneumonia was not significantly different between antibiotic-resistant and antibiotic-susceptible groups. Mortality rates among patients with pneumococcal pneumonia caused by penicillin-, cephalosporin-, or macrolide-resistant strains were not higher than those with antibiotic-susceptible pneumococcal pneumonia. Bacteremia and mechanical ventilation were significant risk factors for death, but any kind of antibiotic resistance was not associated with increased mortality due to pneumococcal pneumonia. Outcome of pneumococcal pneumonia was not significantly affected by drug resistance, and current antimicrobial regimens are mostly effective in the treatment of pneumococcal pneumonia, despite the widespread emergence of in vitro resistance.

Epidemiological typing of Streptococcus pneumoniae from various sources in Sweden and India using Box A PCR fingerprinting.

Epidemiological typing of Streptococcus pneumoniae is essential to determine strain relatedness and also to trace resistant clones. The novel Box A PCR assay was used for characterization of S. pneumoniae isolates from two Scandinavian countries and to compare those from India on the Asian continent. In addition, the assay was employed to determine the clonality of 25 pneumococcal strains from an outbreak in a day-care centre in Linköping, Sweden. All 25 showed a unique pattern with 100% homology for 24 of them, thereby establishing the clonal nature of the outbreak. The pneumococcal strains involved in the outbreak belonged to serotype 9 and were resistant to penicillin, with an MIC value of 2 mg/l. Thirty-eight genotypes were obtained when the Box A results were analysed by computer (Molecular Analyst Software with GelCompar). The discriminatory index of the method was D=0.98, which indicates excellent performance. No major segregation of strains from the different geographical locations was observed when a lower level of similarity was used for typing (80%, 13 types). However, at the level chosen for genotyping, 95% (38 genotypes) there was a clear geographical segregation. No correlation between genotype and serotype was seen as strains from a common place of origin were most often of different serotypes. Computer-assisted analysis of the results of Box A PCR typing facilitated the evaluation.

Fluoroquinolone resistance in clinical isolates of Streptococcus pneumoniae from Asian countries: ANSORP study.

Seventeen clinical isolates of Streptococcus pneumoniae showing reduced susceptibility to ciprofloxacin (MIC >/= 4 micro g/ml) collected from eight different Asian countries were analyzed by antimicrobial susceptibility, serotyping, pulsed-field gel electrophoresis (PFGE), and DNA sequencing of the quinolone resistance-determining regions (QRDRs) in gyrA, gyrB, parC, and parE. All isolates but one showed more than one amino acid alteration in QRDRs of four responsible genes. Ile460 --> Val in parE was the most common mutation. Data suggest that Lys137 --> Asn in parC may be a primary step in the development of high-level and multiple FQ resistance. An additional mutation of Ser81 --> Phe in gyrA resulted in high-level resistance to ciprofloxacin, levofloxacin, and gatifloxacin, whereas Ser79 --> Phe in parC may exert an important role in the development of moxifloxacin resistance. Two novel amino acid changes in gyrB, Ala390 --> Val and Asn423 --> Thr, were found. Data from PFGE suggest an introduction and local spread of multiple resistant Spain(23F)-1 clone in Hong Kong, but isolates from other Asian countries were not related to this clone.

Brain abscess and other protean manifestations of actinomycosis.

Reports on human actinomycosis documented by culture have been infrequent, particularly from India. The present paper reports 12 cases of actinomycosis confirmed by culture. These include two unusual cases of actinomycosis of the brain with no granules in the pus, two pulmonary cases, one renal, one abdominal, and six cervico-facial. The lethal outcome with organ involvement, and the chronicity in some cases, warrant early etiologic confirmation and prompt treatment.