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• Identification of genetic recombination between Acinetobacter species based on multilocus sequence analysis.

  Authors: Dae Hun Kim, Young Kyoung Park, Ji Young Choi, Kwan Soo Ko

  Diagnostic microbiology and infectious disease.

  During multilocus sequence analysis of Acinetobacter calcoaceticus-Acinetobacter baumannii complex, we identified the evidence of recent genetic recombination between 2 Acinetobacter species. While 3During multilocus sequence analysis of Acinetobacter calcoaceticus-Acinetobacter baumannii complex, we identified the evidence of recent genetic recombination between 2 Acinetobacter species. While 3 isolates belonged to A. nosocomialis based on 16S rRNA, gyrB, fusA, gdhB, and rplB gene sequences, they showed close relationships with Acinetobacter genomic species 'close to 13TU' in rpoB, recA, cpn60, rpoD, and gltA gene trees.

• Multidrug resistance among Acinetobacter spp. in the USA and activity profile of key agents: results from CAPITAL Surveillance 2010.

  Authors: Anne Marie Queenan, Chris M Pillar, Jennifer Deane, Daniel F Sahm, A Simon Lynch, Robert K Flamm, Janet Peterson, Todd A Davies

  Diagnostic microbiology and infectious disease.

  Multidrug resistance among Acinetobacter spp. leaves few effective antibiotic options for treatment. To monitor antibiotic resistance in Acinetobacter spp., the US CAPITAL 2010 Surveillance data wereMultidrug resistance among Acinetobacter spp. leaves few effective antibiotic options for treatment. To monitor antibiotic resistance in Acinetobacter spp., the US CAPITAL 2010 Surveillance data were evaluated by patient demographics, specimen source, and hospital ward. Isolates (N=514) were collected from 65 sites across the USA and Puerto Rico. Isolates were centrally tested for susceptibility to carbapenems and key antimicrobials by broth microdilution. Colistin was the most effective agent tested, with 95% susceptibility. The overall susceptibility of Acinetobacter spp. was low (39% for piperacillin/tazobactam, 41% for levofloxacin, 45% for ceftazidime, 47-51% for the carbapenems, and 58% for tobramycin). Multidrug resistance (MDR), defined as resistance to ≥3 antimicrobial agent groups, was detected in 54% of the isolates. MDR isolates were most common among elderly patients (65%), lower respiratory tract isolates (62%), and inpatient/intensive care unit isolates (54-58%). These data update trends in the distribution and prevalence of the MDR phenotype in Acinetobacter spp.

• Assessment of the mosaic structure in the Helicobacter pylori cagA gene 3'-region using an improved polymerase chain reaction-based assay.

  Authors: Hans-Jürg Monstein, Anna Ryberg, Anneli Karlsson

  Diagnostic microbiology and infectious disease.

  The mosaic structure of the cagA gene has been suggested to affect Helicobacter pylori CagA-associated pathogenesis. An improved polymerase chain reaction assay allowed for a rapid and detailedThe mosaic structure of the cagA gene has been suggested to affect Helicobacter pylori CagA-associated pathogenesis. An improved polymerase chain reaction assay allowed for a rapid and detailed molecular analysis of the cagA gene 3'-region in a single amplification step, followed by amplicon sequencing using universal M13 and T7 sequencing primers.

• Activity of telavancin compared to other agents against coagulase-negative staphylococci with different resistotypes by time kill.

  Authors: Gengrong Lin, Glenn A Pankuch, Peter C Appelbaum, Klaudia Kosowska-Shick

  Diagnostic microbiology and infectious disease.

  Among 10 coagulase-negative staphylococci, telavancin, quinupristin/dalfopristin, and tigecycline were the most potent antimicrobials. Telavancin exhibited bactericidal effect to 9 strains out of 10Among 10 coagulase-negative staphylococci, telavancin, quinupristin/dalfopristin, and tigecycline were the most potent antimicrobials. Telavancin exhibited bactericidal effect to 9 strains out of 10 tested at 4× MIC after 24 h of exposure similar to those of vancomycin and daptomycin. By contrast, linezolid was mainly bacteriostatic and teicoplanin was bactericidal to 7 strains tested at 4× MIC after 24 h.

• Comparison of five diagnostic modalities for direct detection of Borrelia burgdorferi in patients with early Lyme disease.

  Authors: Dionysios Liveris, Ira Schwartz, Donna McKenna, John Nowakowski, Robert Nadelman, Joseph Demarco, Radha Iyer, Susan Bittker, Denise Cooper, Diane Holmgren, Gary P Wormser

  Diagnostic microbiology and infectious disease.

  Lyme disease, the most commonly reported tick-borne infection in North America, is caused by infection with the spirochete Borrelia burgdorferi. Although an accurate clinical diagnosis can often beLyme disease, the most commonly reported tick-borne infection in North America, is caused by infection with the spirochete Borrelia burgdorferi. Although an accurate clinical diagnosis can often be made based on the presence of erythema migrans, in research studies microbiologic or molecular microbiologic confirmation of the diagnosis may be required. In this study, we evaluated the sensitivity of 5 direct diagnostic methods (culture and nested polymerase chain reaction [PCR] of a 2-mm skin biopsy specimen, nested PCR and quantitative PCR (qPCR) performed on the same 1-mL aliquot of plasma and a novel qPCR-blood culture method) in 66 untreated adult patients with erythema migrans. Results of one or more of these tests were positive in 93.9% of the patients. Culture was more sensitive than PCR for both skin and blood, but the difference was only statistically significant for blood samples (P<0.005). Blood culture was significantly more likely to be positive in patients with multiple erythema migrans skin lesions compared to those with a single lesion (P=0.001). Positive test results among the 48 patients for whom all 5 assays were performed invariably included either a positive blood or a skin culture. The results of this study demonstrate that direct detection methods such as PCR and culture are highly sensitive in untreated adult patients with erythema migrans. This enabled microbiologic or molecular microbiologic confirmation of the diagnosis of B. burgdorferi infection in all but 4 (6.1%) of the 66 patients evaluated.

• Evaluation of commercial screening tests and blot assays for the diagnosis of Lyme borreliosis.

  Authors: Laurent Busson, Marijke Reynders, Sigi Van den Wijngaert, Hafid Dahma, Marc Decolvenaer, Liesbet Vasseur, Olivier Vandenberg

  Diagnostic microbiology and infectious disease.

  The performance of 4 screening tests and 10 blot assays for the serologic diagnosis of Lyme borreliosis in a Belgian population was evaluated. A total of 196 sera were tested: 36 Lyme borreliosis atThe performance of 4 screening tests and 10 blot assays for the serologic diagnosis of Lyme borreliosis in a Belgian population was evaluated. A total of 196 sera were tested: 36 Lyme borreliosis at different stages of the disease, 50 healthy blood donors, and 110 representing various clinical circumstances. The DiaSorin Liaison and Euroimmun Anti-Borrelia screening tests were evaluated. The tested blot assays were Virotech Borrelia LINE tests WE222, WE225, and WE224, as well as Mikrogen recomLine Borrelia and Viramed ViraStripe. The specificity of IgG was acceptable for the different assays. For IgM, DiaSorin Liaison Borrelia IgM Quant, Mikrogen recomLine, and Viramed ViraStripe lacked specificity. Interestingly, a higher rate of falsely reactive samples was observed in the group of patients suffering from malaria. Serological diagnosis of Lyme borreliosis remains challenging; assays should be evaluated in the population where they are intended to be used.

• Molecular characterization of fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates from Shanghai, China.

  Authors: Cuiyun Zhu, Yongxin Zhang, Yinzhong Shen, Gilman Kit Hang Siu, Wenjuan Wu, Xueqin Qian, Guilin Deng, Yan Xu, Ricky Lau, Xiaoyong Fan, Wenhong Zhang, Hongzhou Lu, Wing-Cheong Yam

  Diagnostic microbiology and infectious disease.

  China is one of the countries with the highest prevalence of fluoroquinolone-resistant (FQ(r)) Mycobacterium tuberculosis. Nevertheless, knowledge on the molecular characterization of the FQ(r)M.China is one of the countries with the highest prevalence of fluoroquinolone-resistant (FQ(r)) Mycobacterium tuberculosis. Nevertheless, knowledge on the molecular characterization of the FQ(r)M. tuberculosis strains of this region remains very limited. This study was performed to investigate the frequencies and types of mutations present in FQ(r)M. tuberculosis clinical isolates collected in Shanghai, China. A total of 206 FQ(r)M. tuberculosis strains and 21 ofloxacin-sensitive (FQ(s)) M. tuberculosis strains were isolated from patients with pulmonary tuberculosis in Shanghai. The phenotypic drug susceptibilities were determined by the proportion method, and the mutations inside quinolone resistance-determining region (QRDR) of gyrA and gyrB genes were identified by DNA sequence analyses. Among 206 FQ(r)M. tuberculosis strains, 44% (90/206) were multidrug-resistant isolates and 39% (81/206) were extensively drug-resistant isolates. Only 9% (19/206) were monoresistant to ofloxacin. In total, 79.1% (163/206) of FQ(r) isolates harboured mutations in either gyrA or gyrB QRDR. Mutations in gyrA QRDR were found in 75.7% (156/206) of FQ(r) clinical isolates. Among those gyrA mutants, a majority (75.6%) harboured mutations at amino acid position 94, with D94G being the most frequent amino acid substitution. Mutations in gyrA QRDR showed 100% positive predictive value for FQ(r)M. tuberculosis in China. Mutations in gyrB were observed in 15.5% (32/206) of FQ(r) clinical isolates. Ten novel mutations were identified in gyrB. However, most of them also harboured mutations in gyrA, limiting their contribution to FQ(r) resistance in M. tuberculosis. Our findings indicated that, similar to other geographic regions, mutations in gyrA were shown to be the major mechanism of FQ(r) resistance in M. tuberculosis isolates. The mutations in gyrA QRDR can be a good molecular surrogate marker for detecting FQ(r)M. tuberculosis in China.

• The preanalytical optimization of blood cultures: a review and the clinical importance of benchmarking in 5 Belgian hospitals.

  Authors: Elise Willems, Annick Smismans, Reinoud Cartuyvels, Guy Coppens, Kristien Van Vaerenbergh, Anne-Marie Van den Abeele, Johan Frans

  Diagnostic microbiology and infectious disease. 73(1):1-8.

  Bloodstream infections remain a major challenge in medicine. Optimal detection of pathogens is only possible if the quality of preanalytical factors is thoroughly controlled. Since the laboratory isBloodstream infections remain a major challenge in medicine. Optimal detection of pathogens is only possible if the quality of preanalytical factors is thoroughly controlled. Since the laboratory is responsible for this preanalytical phase, the quality control of critical factors should be integrated in its quality control program. The numerous recommendations regarding blood culture collection contain controversies. Only an unambiguous guideline permits standardization and interlaboratory quality control. We present an evidence-based concise guideline of critical preanalytical determinants for blood culture collection and summarize key performance indicators with their concomitant target values. In an attempt to benchmark, we compared the true-positive rate, contamination rate, and collected blood volume of blood culture bottles in 5 Belgian hospital laboratories. The true-positive blood culture rate fell within previously defined acceptation criteria by Baron et al. (2005) in all 5 hospitals, whereas the contamination rate exceeded the target value in 4 locations. Most unexpected, in each of the 5 laboratories, more than one third of the blood culture bottles were incorrectly filled, irrespective of the manufacturer of the blood culture vials. As a consequence of this shortcoming, one manufacturer recently developed an automatic blood volume monitoring system. In conclusion, clear recommendations for standardized blood culture collection combined with quality control of critical factors of the preanalytical phase are essential for diagnostic blood culture improvement.

• Comparison of three preparatory methods for detection of bacteremia by MALDI-TOF mass spectrometry.

  Authors: Ryan T Saffert, Scott A Cunningham, Jayawant Mandrekar, Robin Patel

  Diagnostic microbiology and infectious disease. 73(1):21-6.

  We evaluated 3 preparatory methods for processing positive blood culture bottle broths prior to matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)We evaluated 3 preparatory methods for processing positive blood culture bottle broths prior to matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis-differential centrifugation, 10% sodium dodecyl sulfate (SDS), and the Sepsityper kit. Initial evaluation used genus and species level cutoff scores of 1.700-1.999 and ≥2.000, respectively. Processing of an initial 79 blood bottle cultures by differential centrifugation yielded correct identifications of 51 (65%) to the genus and 34 (43%) to the species levels. One hundred and one different blood bottles were simultaneously processed with 10% SDS and Sepsityper. Both yielded genus-level identifications for 77 (76%), and the 2 yielded species-level identifications for 49 (49%) and 54 (54%) of bottles, respectively. Adjustment of the score cutoff criteria for genus (1.500-1.699) and species (≥1.700) improved identification percentages, particularly for species-level identifications (P < 0.05).

• Genetic variants of the MBL2 gene are associated with mortality in pneumococcal sepsis.

  Authors: José Garnacho-Montero, Emilio García-Cabrera, Rocio Jiménez-Álvarez, Ana Díaz-Martín, Jaume Revuelto-Rey, Javier Aznar-Martín, Carmen Garnacho-Montero

  Diagnostic microbiology and infectious disease. 73(1):39-44.

  Studies evaluating associations between polymorphisms of innate immunity genes and prognosis of infectious diseases have yielded conflicting results. Our aim was to assess the impact on mortality ofStudies evaluating associations between polymorphisms of innate immunity genes and prognosis of infectious diseases have yielded conflicting results. Our aim was to assess the impact on mortality of different genotypic variants of the innate immunity in patients with pneumococcal sepsis. All adults admitted to the hospital with diagnosis of sepsis caused by Streptococcus pneumoniae were enrolled and single-nucleotide polymorphisms (SNP) in mannose-binding lectin 2 (MBL2), toll-like receptor (TLR) 2, TLR4, and Fcγ receptor IIa genes were genotyped. Underlying diseases, severity of illness, and antibiotic management were also recorded. We included 117 patients: 98 pneumonias (83.6%), 17 meningitis (14.5%), and 2 patients (1.9%) with primary pneumococcal bacteremia. Allelic variants of the MBL2 gene (individuals heterozygous or homozygous for one of the 3 allelic variants B, C, and D: AO/OO) were present in 37 patients (32%), T399I polymorphism in TLR4 in 19 (16.2%), TLR4 D299G/T399I in 11 (9.4%), TLR2 R753Q in 3 (2.5%), and FcγRIIa-R/R131 in 26 patients (23%). Factors associated independently with in-hospital mortality were SNP MBL2 AO/OO (adjusted hazard ratios [aHR] 3.2, 95% confidence interval [CI] 1.01-9.8) and septic shock (aHR 15.3, 95% CI 3.5-36.5), whereas first adequate antibiotic dose ≤4 h was a protective factor (aHR 0.2, 95% CI 0.06-0.8). SNP MBL2 AO/OO (aHR 2.2, 95% CI 1.1-8.1) remained as a variable independently associated with 90-day mortality. In conclusion, variant alleles in the MBL2 gene are independently associated with in-hospital and medium-term mortalities in patients admitted to the hospital with pneumococcal sepsis.

• Evaluation of the MALDI TOF-MS method for identification of Candida strains isolated from blood cultures.

  Authors: Görkem Yaman, Işın Akyar, Simge Can

  Diagnostic microbiology and infectious disease. 73(1):65-7.

  Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF-MS) allows rapid and accurate identification of microorganisms. It is being used increasingly and becoming anMatrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF-MS) allows rapid and accurate identification of microorganisms. It is being used increasingly and becoming an important tool in clinical laboratories. Phenotypic identification of Candida species remains labor- and time consuming, and the results may sometimes be inconclusive. Rapid and reliable species identification of Candida is essential for antifungal treatment due to species-specific susceptibility patterns. In this study, we evaluated the performance of MALDI TOF-MS for identification of Candida strains. A total of 281 clinical Candida strains isolated from blood cultures were included in this study. Candida species were identified with conventional methods and automated VITEK 2 YST panels as well as with MALDI TOF-MS. Isolates with discrepant results were subjected to DNA sequencing of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2). Ninety-four percent of the isolates were identified correctly by VITEK 2 and MALDI TOF-MS. Altogether, MALDI-TOF MS yielded the correct species identification for 281 (100%) clinical Candida isolates. MALDI-TOF proved to be a rapid and reliable method for identification of Candida strains in the clinical laboratory.

• Spontaneous bacterial empyema due to non-O1, non-O139 Vibrio cholerae in a cirrhotic patient with hepatocellular carcinoma.

  Authors: Chih-Cheng Lai, Wei-Lun Liu, Yu-Hsin Chiu, Shiow-Jen Gau, Po-Ren Hsueh

  Diagnostic microbiology and infectious disease. 73(1):84-5.

  Vibrio cholerae is known as a common etiology of epidemic diarrheal disease and rarely causes extra-intestinal infections. In this report, we described a cirrhotic patient with hepatocellularVibrio cholerae is known as a common etiology of epidemic diarrheal disease and rarely causes extra-intestinal infections. In this report, we described a cirrhotic patient with hepatocellular carcinoma who developed spontaneous bacterial empyema due to non-O1, non-O139 V. cholerae. The patient was successfully treated with antimicrobial agents and percutaneous drainage.

• Influenza viral load and rapid influenza diagnostic tests in children and adults.

  Authors: Michimaru Hara, Masao Morihara, Shinichi Takao, Shinji Fukuda, Yukie Shimazu, Yukie Tanizawa, Takeshi Matsuo

  Diagnostic microbiology and infectious disease. 73(1):99-100.

  We compared children and adults with regard to rapid influenza test sensitivity and viral load. Specimen volumes were measured, rapid tests were conducted, and viral load was determined. There was noWe compared children and adults with regard to rapid influenza test sensitivity and viral load. Specimen volumes were measured, rapid tests were conducted, and viral load was determined. There was no difference between children and adults in test sensitivity or viral load, but children had higher specimen volumes.

• Lack of evidence for rapid transmission of Lyme disease following a tick bite.

  Authors: Matthew J Binnicker, Elitza S Theel, Bobbi S Pritt

  Diagnostic microbiology and infectious disease. 73(1):102-3.

• Can mailed swab samples be dry-shipped for the detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis by nucleic acid amplification tests?

  Authors: Charlotte A Gaydos, Carol Farshy, Mathilda Barnes, Nicole Quinn, Patricia Agreda, Charles A Rivers, Jane Schwebke, John Papp

  Diagnostic microbiology and infectious disease. 73(1):16-20.

  Dry-shipped and mailed vaginal swabs collected at home have been used in research studies for the detection of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC), and Trichomonas vaginalis (TV)Dry-shipped and mailed vaginal swabs collected at home have been used in research studies for the detection of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC), and Trichomonas vaginalis (TV) by nucleic acid amplification tests (NAATs) in screening programs. A verification study was performed to compare the limit of detection of CT, GC, and TV on swabs that were dry-shipped to paired swabs that were wet-shipped in transport media through the US mail. The Centers for Disease Control and Prevention prepared inocula in sterile water to mock simulated urogenital swabs with high to low concentrations of CT and GC. Replicate swabs were inoculated with 100 μL of dilutions and were dry transported or placed into commercial transport media ("wet") for mailing for NAAT testing. The University of Alabama prepared replicate concentrations of TV, which were similarly shipped and tested by NAAT. All paired dry and wet swabs were detectable for CT. For GC, all paired dry and wet swabs were detectable for GC at concentrations ≥10(3). At 10(2) and 10 CFU/mL, the 10 replicate GC results were variably positive. For TV, wet and dry shipped concentrations >10(2) TV/mL tested positive, while results at 10 TV/mL were negative for dry swabs. Holding replicate dry swabs at 55 (○)C 5 days before testing did not affect results. NAATs were able to detect CT, GC, and TV on dry transported swabs. Using NAATs for testing home-collected, urogenital swabs mailed in a dry state to a laboratory may be useful for outreach screening programs.

• First countrywide survey of third-generation cephalosporin-resistant Escherichia coli from broilers, swine, and cattle in Switzerland.

  Authors: Andrea Endimiani, Alexandra Rossano, Daniel Kunz, Gudrun Overesch, Vincent Perreten

  Diagnostic microbiology and infectious disease. 73(1):31-8.

  The herd prevalence of third-generation cephalosporin-resistant Escherichia coli (3GC-R-Ec) was determined for broilers (25.0% [95% confidence interval (CI) 17.6-33.7%]), pigs (3.3% [(95% CIThe herd prevalence of third-generation cephalosporin-resistant Escherichia coli (3GC-R-Ec) was determined for broilers (25.0% [95% confidence interval (CI) 17.6-33.7%]), pigs (3.3% [(95% CI 0.4-11.5%]), and cattle (3.9% [95% CI 0.5-13.5%]), using a sampling strategy that was representative of the livestock population slaughtered in Switzerland between October 2010 and April 2011. The 3GC-R-Ec isolates were characterized by the measurement of the MICs of various antibiotics, microarray analyses, analytical isoelectric focusing, polymerase chain reaction and DNA sequencing for bla genes, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. CMY-2 (n = 12), CTX-M-1 (n = 11), SHV-12 (n = 5), TEM-52 (n = 3), CTX-M-15 (n = 2), and CTX-M-3 (n = 1) producers were found. The majority of CMY-2 producers fell into 1 PFGE cluster, which predominantly contained ST61, whereas the CTX-M types were carried by heterogeneous clones of E. coli, as shown by the numerous PFGE profiles and STs that were found. This is the first national Swiss study that focuses on the spread of 3GC-R Enterobacteriaceae among slaughtered animals.

• The changing epidemiology of healthcare-associated candidemia over three decades.

  Authors: Daniel Diekema, Sophie Arbefeville, Linda Boyken, Jennifer Kroeger, Michael Pfaller

  Diagnostic microbiology and infectious disease. 73(1):45-8.

  We describe the epidemiology of healthcare-associated candidemia (HAC) in our tertiary care hospital, in comparison with both the pre-fluconazole (pre-FLU) and pre-echinocandin (pre-EC) eras. WeWe describe the epidemiology of healthcare-associated candidemia (HAC) in our tertiary care hospital, in comparison with both the pre-fluconazole (pre-FLU) and pre-echinocandin (pre-EC) eras. We identified all patients with HAC using microbiology records from 1/2004 to 12/2007, reviewed medical records, and pulled isolates for testing. We compared mortality, underlying illness, Candida species distribution, and antifungal susceptibility with 2 prior University of Iowa cohorts (88 patients from 1983 to 1986 [pre-FLU], and 108 from 1997 to 2001 [pre-EC]). Of 108 patients with HAC from 2004 to 2007, species distribution was 47% C. albicans, 29% C. glabrata, 12% C. parapsilosis, 6% C. tropicalis, and no C. krusei. Compared with pre-FLU and pre-EC eras, there was a reduction in % C. albicans (from 61% and 60%, respectively), an increase in % C. glabrata (from 0% and 16%), and no change in % C. parapsilosis over time (12% and 12%). In-hospital mortality was lower in 2004-2007 than both pre-FLU and pre-EC (31% versus 57-61%), and 30-day mortality was also lower (33% versus 48% in pre-EC). Mean Charlson index was lower for the 2004-2007 cohort than pre-EC (3.0 versus 3.4)-fewer patients had leukemia or lymphoma (8% versus 16%) or other malignancies (18% versus 24%), while more were surgical patients (58% versus 48%). Using the new Clinical and Laboratory Standards Institute breakpoints for FLU and caspofungin, we found no caspofungin resistance, and FLU resistance only among C. glabrata (15% had FLU MICs >32 µg/mL). The epidemiology of HAC is changing at our hospital, with continued emergence of C. glabrata, fewer cases among oncology patients, and lower in-hospital and 30-day mortality.

• Role of quantitative CSF microscopy to predict culture status and outcome in HIV-associated cryptococcal meningitis in a Brazilian cohort.

  Authors: José E Vidal, Juliana Gerhardt, Erique J Peixoto de Miranda, Rafi F Dauar, Gilberto S Oliveira Filho, Augusto C Penalva de Oliveira, David R Boulware

  Diagnostic microbiology and infectious disease. 73(1):68-73.

  This retrospective study aimed to evaluate the clinical, laboratory, and quantitative cerebrospinal fluid (CSF) cryptococcal cell counts for associations with in-hospital outcomes of HIV-infectedThis retrospective study aimed to evaluate the clinical, laboratory, and quantitative cerebrospinal fluid (CSF) cryptococcal cell counts for associations with in-hospital outcomes of HIV-infected patients with cryptococcal meningitis. Ninety-eight HIV-infected adult patients with CSF culture-proven cryptococcal meningitis were admitted between January 2006 and June 2008 at a referral center in Sao Paulo, Brazil. Cryptococcal meningitis was the first AIDS-defining illness in 69%, of whom 97% (95/98) had known prior HIV infection. The median CD4+ T-cell count was 39 cells/μL (interquartile range 17-87 cells/μL). Prior antiretroviral therapy was reported in 50%. Failure to sterilize the CSF by 7-14 days was associated with baseline fungal burden of ≥10 yeasts/μL by quantitative CSF microscopy (odds ratio [OR] = 15.3, 95% confidence interval [CI] 4.1-56.7; P < 0.001) and positive blood cultures (OR = 11.5, 95% CI 1.2-109; P = 0.034). At 7-14 days, ≥10 yeasts/μL CSF was associated with positive CSF cultures in 98% versus 36% with <10 yeasts/μL CSF (P < 0.001). In-hospital mortality was 30% and was associated with symptoms duration for >14 days, altered mental status (P < 0.001), CSF white blood cell counts <5 cells/μL (P = 0.027), intracranial hypertension (P = 0.011), viral loads >50,000 copies/mL (P = 0.036), ≥10 yeasts/μL CSF at 7-14 days (P = 0.038), and intracranial pressure >50 cmH(2)0 at 7-14 days (P = 0.007). In conclusion, most patients were aware of their HIV status. Fungal burden of ≥10 yeasts/μL by quantitative CSF microscopy predicted current CSF culture status and may be useful to customize the induction therapy. High uncontrolled intracranial pressure was associated with mortality.

• Two hundred and eleven cases of Candida osteomyelitis: 17 case reports and a review of the literature.

  Authors: Amy K Slenker, Scott W Keith, David L Horn

  Diagnostic microbiology and infectious disease. 73(1):89-93.

  Candida osteomyelitis is a rare infection. We present 17 cases of Candida osteomyelitis from our institution and review 194 patients from the literature. The median age of patients was 57 ± 22 yearsCandida osteomyelitis is a rare infection. We present 17 cases of Candida osteomyelitis from our institution and review 194 patients from the literature. The median age of patients was 57 ± 22 years (range, 18-90 years) with 68% male. Comorbidities associated with this infection include prior surgery (62%), broad-spectrum antibiotics (40%), central venous catheter insertion (19%), and immunosuppression (17%). The most common infecting species were Candida albicans (69%), Candida tropicalis (15%), and Candida glabrata (8%). Most initial antifungal regimens included amphotericin B (59%); however, fluconazole is increasingly being utilized for treatment of this infection (26%). Echinocandins were used infrequently (4%). Median length of treatment was 3 ± 4.5 months (mean, 4.2 months; range, 18 days to 36 months). The overall success rate of therapy was 91%, with 75% of patients cured by 6 months. The crude mortality rate was 12% with an attributable mortality rate of 6%.

• Reply to the apropos "Evaluation of third-generation RIDASCREEN enzyme immunoassay for the detection of norovirus antigens in stool samples of hospitalized children in Belém, Pará, Brazil".

  Authors: Jones Anderson Monteiro Siqueira, Alexandre C Linhares, Yvone Benchimol Gabbay

  Diagnostic microbiology and infectious disease. 73(1):101-2.

• Clinical evidence for rapid transmission of Lyme disease following a tickbite: response to Piesman and Gray.

  Authors: Raphael B Stricker, Eleanor D Hynote, Phyllis C Mervine

  Diagnostic microbiology and infectious disease. 73(1):104-5.

• Detection of KPC-2 and qnrS1 in clinical isolates of Morganella morganii from China.

  Authors: Jia Chang Cai, Wei Yang, Yan Yan Hu, Rong Zhang, Hong Wei Zhou, Gong-Xiang Chen

  Diagnostic microbiology and infectious disease.

  Seven carbapenem-nonsusceptible Morganella morganii isolates, which have similar antibiotic susceptibility profiles, were isolated over a 5-month period. MICs of imipenem, meropenem, and ertapenemSeven carbapenem-nonsusceptible Morganella morganii isolates, which have similar antibiotic susceptibility profiles, were isolated over a 5-month period. MICs of imipenem, meropenem, and ertapenem were 8, 1, and 0.25 to 0.5 μg/mL, respectively. Pulsed-field gel electrophoresis indicated that 6 isolates were indistinguishable or closely related. Carbapenem resistance can be transferred from M. morganii to Escherichia coli by conjugation. All M. morganii isolates and E. coli transconjugants produced KPC-2 and carried the qnrS1 gene. Production of KPC-2 mainly contributed to the carbapenem resistance in M. morganii. KPC-2-producing M. morganii clonally spread in a hospital in China.

• Kinetics of CMV seroconversion in a Swiss pregnant women population.

  Authors: Gregory T Maine, René Stricker, Reto Stricker

  Diagnostic microbiology and infectious disease.

  Retrospective evaluation of the kinetics of cytomegalovirus (CMV) seroconversion with CMV IgM, IgG, and IgG avidity assays, in a Swiss pregnant women population, has shown that the current publishedRetrospective evaluation of the kinetics of cytomegalovirus (CMV) seroconversion with CMV IgM, IgG, and IgG avidity assays, in a Swiss pregnant women population, has shown that the current published CMV serologic diagnostic algorithms were valid and fit for use. In 19% of the cases analyzed, CMV-specific IgM was detected before IgG.

• Isolation of a capnophilic Escherichia coli strain from an empyemic patient.

  Authors: Weiping Lu, Kai Chang, Shaoli Deng, Min Li, Junji Wang, Ji Xia, Hengliu Huang, Ming Chen

  Diagnostic microbiology and infectious disease.

• Evaluation of BACTEC Plus aerobic and anaerobic blood culture bottles and BacT/Alert FAN aerobic and anaerobic blood culture bottles for the detection of bacteremia in ICU patients.

  Authors: Kyoung Ho Roh, Ju Yeon Kim, Ha Nui Kim, Hye Jin Lee, Jang Wook Sohn, Min Ja Kim, Yunjung Cho, Young Kee Kim, Chang Kyu Lee

  Diagnostic microbiology and infectious disease.

  Blood culture is the most valuable laboratory test for the diagnosis of bacteremia and sepsis. The BACTEC FX and BacT/Alert 3D automated blood culture systems are commonly used in Korean health careBlood culture is the most valuable laboratory test for the diagnosis of bacteremia and sepsis. The BACTEC FX and BacT/Alert 3D automated blood culture systems are commonly used in Korean health care facilities. A controlled clinical evaluation of the resin-containing BACTEC Plus aerobic (BA) and anaerobic (BN), and the charcoal-containing FAN aerobic (FA) and anaerobic (FN) bottles using blood from intensive care unit (ICU) patients was designed. The performances of these 2 systems with media containing particle absorbing antimicrobial agents were evaluated using the culture positivity rate and time to detection (TTD). TTD was collected using data management systems, either the Epicenter (BD Diagnostic Systems) or the hospital laboratory information system. A total of 1539 four-bottle sets were collected from 270 patients in medical and surgical ICUs. Blood culture samples included 1539 bottles each of BA, BN, FA, and FN, and yielded 113 (7.3%), 90 (5.8%), 104 (6.8%), and 80 (5.2%) positive bacterial or fungal isolates, respectively. There were significant differences between the resin-containing BA and BN samples in culture positivity and also between the charcoal-containing FA and FN samples, especially for Escherichia coli (25/27 versus 17/27, P < 0.05) and Acinetobacter baumannii (14/15 versus 7/15, P < 0.05). Significantly shorter recovery time was observed in BACTEC Plus aerobic bottles than in FAN aerobic bottles (17.2 and 24.7 h, respectively) (P < 0.001).

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