Diagnostic microbiology and infectious disease

Publications in this journal

• Article: Rapid method for Mycobacterium tuberculosis identification using electrospray ionization tandem mass spectrometry analysis of mycolic acids.

  Rafał Szewczyk, Konrad Kowalski, Beata Janiszewska-Drobinska, Magdalena Druszczyńska
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  ABSTRACT: Mycolic acids (MAs), which play a crucial role in the architecture of mycobacterial cell walls, were analyzed using electrospray ionization tandem mass spectrometry. A targeted analysis based on the 10 most abundant and characteristic multiple reaction monitoring pairs was used to profile the crude fatty acid mixtures from Mtb and several nontuberculous mycobacterial strains. Comparative analysis yielded unique profiles for MAs, enabling the reliable identification of mycobacterial species. In a case-control study of tuberculosis (TB) and non-TB Polish patients, we demonstrated the potential diagnostic utility of our approach for the rapid diagnosis of active TB with sensitivity and specificity surpassing those of existing methods. This robust method allows the identification of TB-positive patients after 2 h of sample preparation in the case of direct sputum analysis or 10 days of culturing, both of which are followed by 1 min of liquid chromatography-tandem mass spectrometry analysis.
  Diagnostic microbiology and infectious disease 05/2013;
• Article: Trends in Klebsiella pneumoniae carbapenemase-positive K. pneumoniae in US hospitals: report from the 2007-2009 SENTRY Antimicrobial Surveillance Program.

  Robyn M Kaiser, Mariana Castanheira, Ronald N Jones, Fred Tenover, Ruth Lynfield
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  ABSTRACT: We report the prevalence of carbapenemase-positive Klebsiella pneumoniae among clinical isolates collected from US medical centers (n = 42) from 2007-2009 through the SENTRY Antimicrobial Surveillance Program. Isolates with imipenem or meropenem MIC ≥2 μg/mL were screened by PCR for various carbapenemase genes. Of 2049 K. pneumoniae isolates, 126 (6.1%) were non-susceptible to imipenem or meropenem. blaKPC was identified in 113 isolates (5.5%). No other carbapenemase genes were identified. For US regions combined, prevalence of K. pneumoniae carbapenemase (KPC)-positive isolates were 5.9% in 2007, 4.9% in 2008, and 5.7% in 2009. Rates were highest in the Mid-Atlantic region (28.6% overall), with fluctuation over time (29%, 23%, and 33% from 2007-2009), followed by the East North Central region (2.4% overall), with a slightly increasing trend (nil, 3.1%, 3.8% from 2007-2009). All KPC-positive organisms were carbapenem non-susceptible according to updated CLSI breakpoints, although all but one was similarly classified according to previous breakpoints.
  Diagnostic microbiology and infectious disease 05/2013;
• Article: Evaluation of interferon-γ release assay in the diagnosis of osteoarticular tuberculosis.

  Hongyan Jia, Liping Pan, Shibing Qin, Fei Liu, Fengjiao Du, Tinglong Lan, Xia Zhang, Rongrong Wei, Boping Du, Zhongquan Liu, Hairong Huang, Zongde Zhang
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  ABSTRACT: The aim of this study was to assess the value of interferon-γ (IFN-γ) release assay (IGRA) (T-SPOT.TB) for patients with suspected osteoarticular tuberculosis (TB) in comparison with conventional and molecular methods. Of 145 patients with suspected osteoarticular TB, recruited from Beijing Chest Hospital between July 2011 and June 2012, 86 (59.3%)had osteoarticular TB (26 with culture-confirmed TB, 60 with probable TB), 24 (16.6%) were not having active TB. The remaining 17 (11.7%) inconclusive TB and 18 (12.4%) possible TB were excluded from final analysis. In addition to conventional tests and molecular method, T-SPOT.TB assay using peripheral blood mononuclear cells to examine IFN-γ response to early secretory antigenic target 6 and culture filtrate protein 10 was also performed. The sensitivity and specificity for T-SPOT.TB assay were 94.2% and 70.8%, respectively. A statistically significant difference in sensitivity was found between T-SPOT.TB assay (94.2%) and other tests (acid-fast bacilli smear (19.7%), culture (34.2%), real-time PCR (36.8%); P < 0.01, respectively). These results suggested that the IGRA assay could provide useful aids in the diagnosis of osteoarticular TB.
  Diagnostic microbiology and infectious disease 05/2013;
• Article: Development and evaluation of multiplex PCR in rapid diagnosis of abdominal tuberculosis.

  Vinaykumar Hallur, Meera Sharma, Sunil Sethi, Kusum Sharma, Abhishek Mewara, Sunil Dhatwalia, Rakesh Yadav, Deepak Bhasin, Saroj Kant Sinha, Satyawati Rana, Kusum Joshi
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  ABSTRACT: The clinical features of abdominal tuberculosis (TB) are non-specific and establishing a diagnosis remains a challenge. A delay in diagnosis is likely to increase the morbidity in these patients. We developed a multiplex polymerase chain reaction (PCR) using 16SrRNA, IS6110, and devR, and evaluated it in comparison with other conventional tests in clinical suspects of abdominal TB. A total of 183 patients with clinical suspicion of abdominal TB (96 patients with intestinal TB and 87 with peritoneal TB) were enrolled for the study. Endoscopic or intraoperative biopsies were collected from patients suspected of intestinal TB and ascitic fluid was collected from patients with a suspicion of peritoneal TB. Of the intestinal tuberculosis group, there were 40 confirmed cases and 56 controls, while of the peritoneal tuberculosis group there were 37 confirmed cases and 50 controls. Multiplex PCR showed a high sensitivity and specificity in both the intestinal TB and peritoneal TB groups. When combined with histopathology, multiplex PCR could detect 97.5% of all the cases in the intestinal tuberculosis group, while in combination adenosine deaminase levels (ADA) in cases of peritoneal tuberculosis it increased the specificity of diagnosis of peritoneal tuberculosis to 95%. In combination with histopathology in suspected intestinal TB cases, and ADA testing in suspected peritoneal TB cases, it can be used as a highly sensitive, specific, and rapid diagnostic tool with the ability to supplement the limitations of other diagnostic modalities.
  Diagnostic microbiology and infectious disease 05/2013; 76(1):51-5.
• Article: Evaluation of 3 different agar media for rapid detection of extended-spectrum β-lactamase-producing Enterobacteriaceae from surveillance samples.

  Elise Willems, Reinoud Cartuyvels, Koen Magerman, Jan Verhaegen
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  ABSTRACT: Rapid detection of extended-spectrum β-lactamase (ESBL)-producing Gram-negative bacilli in surveillance samples of high-risk patients allows early optimization of antimicrobial therapy and timely introduction of infection control procedures. This study evaluated the BLSE (AES Chemunex), chromID ESBL (bioMérieux), and Brilliance ESBL agar (Oxoid) for rapid detection of ESBL-producing Enterobacteriaceae from surveillance samples. A total of 139 perineal and nose samples were processed. Isolated bacterial strains were identified by mass spectrometry. ESBL confirmation was performed by phenotypical and molecular tests. Overall, 16 ESBL-producing Enterobacteriaceae were recovered. The sensitivities after 24 h of incubation were comparable (BLSE, 87.5%; Brilliance ESBL, 87.5%; and chromID, 81.3%). The specificity of chromogenic media (80.7-82.1%) was significantly higher compared to BLSE (60.8%). All 3 media are reliable to screen for ESBL-producing Enterobacteriaceae from surveillance samples. Yet, the main advantages of chromogenic media over BLSE reside in their chromogenic character and higher specificity, reducing the total number of isolates that require further identification and ESBL confirmation testing.
  Diagnostic microbiology and infectious disease 05/2013; 76(1):16-9.
• Article: Hepatitis E virus serological testing in kidney transplant recipients with elevated liver enzymes in 2007-2011 in southeastern France.

  Valérie Moal, Anne Motte, Mamadou Kaba, René Gerolami, Yvon Berland, Philippe Colson
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  ABSTRACT: Hepatitis E virus (HEV) is an emerging cause of acute and chronic hepatitis in Europe, particularly in solid organ transplant recipients. Anti-HEV IgG/IgM testing in kidney transplant recipients with liver biological disturbances indicated high HEV exposure in our geographical area and led to diagnose HEV infection in 6% of cases.
  Diagnostic microbiology and infectious disease 05/2013; 76(1):116-8.
• Article: Phenotypic and genotypic characterization of enterotoxigenic Escherichia coli isolated from U.S. military personnel participating in Operation Bright Star, Egypt, from 2005 to 2009.

  Rania A Nada, Adam Armstrong, Hind I Shaheen, Isabelle Nakhla, John W Sanders, Mark S Riddle, Sylvia Young, Peter Sebeny
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  ABSTRACT: Enterotoxigenic Escherichia coli (ETEC) is a major health problem for travelers to the Middle East. During the autumn months of 2005, 2007, and 2009, U.S. military personnel participated in Operation Bright Star (OBS) exercises in Egypt. Out of 181 military personnel enrolled in a diarrheal surveillance study, E. coli-like colonies were isolated from 170 patients. Isolates were tested for the detection of ETEC enterotoxins and colonization factors (CFs) using phenotypic and genotypic methods. Additionally, we studied the secular trends of ETEC isolates obtained from OBS studies since 1999. ETEC was isolated from 51.2% and 60.0% of the patients based on enzyme-linked immunosorbent assay and polymerase chain reaction (PCR), respectively. Heat stable (ST) was the dominant enterotoxin detected followed by heat labile (LT) and LTST. Additionally, we detected a CF in 59.7% and 67.6% of the ETEC-positive isolates using dot blot and PCR assays, respectively. The predominant CF isolated was CS6 followed by CS3.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Acinetobacter pittii and Acinetobacter nosocomialis among clinical isolates of the Acinetobacter calcoaceticus-baumannii complex in Sichuan, China.

  Xiaohui Wang, Tao Chen, Rujia Yu, Xiaojü Lü, Zhiyong Zong
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  ABSTRACT: Among 82 clinical isolates of the Acinetobacter calcoaceticus-baumannii complex recovered in 13 hospitals of Sichuan, China, in 2011, 13 were Acinetobacter pittii and 2 were Acinetobacter nosocomialis. Multilocus sequence typing revealed a novel sequence type (ST) of A. nosocomialis and 7 novel STs of A. pittii. Most isolates were hospital-acquired and colonized in the respiratory tract, while 6 cases with pneumonia due to A. pittii were identified. This study provided a snapshot of the local incidence of A. pittii and A. nosocomialis.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Identification, clinical aspects, susceptibility pattern, and molecular epidemiology of beta-haemolytic group G Streptococcus anginosus group isolates from central Taiwan.

  Yi-Cheng Chang, Hsueh-Hsia Lo
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  ABSTRACT: No literature is available on the prevalence and clinical aspects of beta-haemolytic group G Streptococcus anginosus group in central Taiwan. In this study, we used 16S rRNA gene sequencing and 16S-23S rDNA intergenic spacer sequencing (where necessary) as the gold standard for molecular identification. Twenty-seven S. anginosus group isolates were identified from 273 beta-haemolytic GGS isolates collected from patients in central Taiwan between February 2007 and August 2011. Of the 27 isolates, 22 were S. anginosus and 5 were Streptococcus constellatus. The 3 commercial methods, Rapid ID 32 Strep, API 20 Strep, and Vitek 2 GP card, identified 77.8%, 40.7%, and 37.0% of S. anginosus group isolates, respectively, with acceptable %ID or probability level. All the S. constellatus isolates possessed the lmb gene (encoding laminin-binding protein); however, none of the S. anginosus isolates possessed this gene. All the 27 isolates were susceptible to penicillin. Five S. anginosus group isolates (18.5%) were resistant to erythromycin. The resistance genes, ermB and mefA, were detected in 3 (2 S. anginosus and 1 S. constellatus) and 2 (2 S. anginosus) isolates, respectively. Pulsed field gel electrophoresis showed that most S. anginosus group isolates were genetically diverse. This is the first study to evaluate 3 commercial methods for the identification of beta-haemolytic group G S. anginosus group species, and only the Rapid ID 32 Strep system showed considerable ability. The clinical aspects, susceptibility pattern, and molecular epidemiology of beta-haemolytic group G S. anginosus group isolates from central Taiwan were also first presented.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Comparison of LED and conventional fluorescence microscopy for detection of acid-fast bacilli in an area with high tuberculosis incidence.

  Manel Marzouk, Asma Ferjani, Mohamed Dhaou, Moufida Haj Ali, Naila Hannachi, Jalel Boukadida
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  ABSTRACT: The objective of the study is to compare the performance of conventional fluorescence microscopy (CFM) and light-emitting diode (LED) fluorescence microscopy (FM) for detection of acid-fast bacilli (AFB) in clinical samples. We included AFB smears, stained using the auramine O method and blindly examined with both CFM and LED-FM. Culture results were used as reference for evaluating the reliability of the FM. We included 180 culture positive specimens and an equal number of culture negative specimens. Sensitivities for the CFM and LED-FM were 79.4% and 82.2%, respectively. Both microscopes had a high specificity (97.2%). The negative-positive (>1 cross) inter-reader agreement of LED-FM and CFM was excellent. Therefore, detection of scanty AFB was higher with LED-FM. Both microscopes were equivalent with respect to time required to read smears. Although it was not faster than CFM, the higher detection of scanty AFB smears combined with ease of use supports the consideration of LED microscopy by all tuberculosis diagnostic laboratories, as a replacement for conventional fluorescence microscopes.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Ciprofloxacin-resistant Escherichia coli isolated from the intestinal microbiota of goats in Greece in the absence of selective pressure.

  George Filioussis, Athanasia Tzivara, Evanthia Petridou, Nektarios D Giadinis, Angeliki R Burriel, Spyridon K Kritas
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  ABSTRACT: The presence of ciprofloxacin resistance commensal Escherichia coli (C-R-Ec) was determined for goats in the absence of selective pressure in Northern and Central Greece. The C-R-Ec was categorized in 3 groups with respect to their phenotypic resistance to other antibiotics as well as the carriage of antibiotic resistance genes. The first group consisted of 7 C-R-Ec that were found also resistant to tetracycline. Among them tet(B) (n = 7), qnr(S) (n = 7), and qnr(B) (n = 3) producers were identified by polymerase chain reaction. The second group consisted of 10 C-R-Ec that were found sensitive to all other antibiotics, and their phenotypic resistance to ciprofloxacin was not attributed to the presence of resistance genes. Finally, the third group consisted of 2 C-R-Ec also resistant to sulfamethoxazole. These strains were not carrying any transferable elements that contribute to resistance either to ciprofloxacin or to sulfamethoxazole. This is the first report of ciprofloxacin-resistant E. coli isolated from goats in Greece.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: High prevalence of multiresistance in levofloxacin-nonsusceptible Streptococcus pneumoniae isolates in Korea.

  Sungkyoung Lee, Seong-Han Kim, Misun Park, Songmee Bae
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  ABSTRACT: Korea exhibits the highest rates of multidrug resistance among Streptococcus pneumoniae. The increasing use of levofloxacin has raised concern about the dissemination of levofloxacin resistance in dominant multidrug-resistant (MDR) clones of our pneumococcal population. A total of 50 levofloxacin-nonsusceptible S. pneumoniae (MIC, ≥4 μg/mL) collected from a multihospital network from 1996 to 2006 were analyzed for serotype, antibiotic resistance profile, quinolone resistance-determining region mutation, and multilocus sequence type. Most levofloxacin-nonsusceptible S. pneumoniae (94.0%) exhibited an MDR phenotype. This phenotype was closely associated with a limited number of epidemic MDR clones that are well-known key agents of the global spread of antimicrobial resistance in S. pneumoniae. However, the clonal dissemination of levofloxacin-nonsusceptible S. pneumoniae was rare. Levofloxacin-nonsusceptible clones with nonvaccine serotypes increased during the post-vaccine era in this study. This result suggests that Korean clinicians must be aware of the levofloxacin resistance trend and need to be more prudent for the first choice of fluoroquinolone for empiric treatment of respiratory tract infections in clinical setting. Moreover, the emergence of new clones and their variations may be more frequently associated with resistance under this selective pressure, such as the introduction of a 7-valent pneumococcal conjugate vaccine into our community.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Comparative in vitro activity of ceftaroline, ceftaroline-avibactam, and other antimicrobial agents against aerobic and anaerobic bacteria cultured from infected diabetic foot wounds.

  Ellie J C Goldstein, Diane M Citron, C Vreni Merriam, Kerin L Tyrrell
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  ABSTRACT: Foot infections are the most common infectious complication of diabetes. Moderate to severe diabetic foot infections (DFI) are typically polymicrobial with both aerobic and anaerobic organisms. The role of MRSA in these wounds has become an increasing concern. To determine if the addition of avibactam, a novel non-beta-lactam beta-lactamase inhibitor, to ceftaroline would be more active than ceftaroline alone, we tested 316 aerobic pathogens and 154 anaerobic recovered from patients with moderate to severe DFI, and compared ceftaroline with and without avibactam to other agents. Testing on aerobes was done by broth microdilution and by agar dilution for anaerobes, according to CLSI M11-A8, and M7-A8 standards. Ceftaroline-avibactam MIC90 for all Staphylococcus spp. including MRSA was 0.5 μg/mL, and for enterococci was 1 μg/mL. The MIC90s for enteric Gram-negative rods was 0.125 μg/mL. The addition of avibactam to ceftaroline reduced the ceftaroline MICs for 2 strains of resistant Enterobacter spp. and for 1 strain of Morganella. Against anaerobic Gram-positive cocci ceftaroline-avibactam had an MIC90 0.125 μg/mL and for clostridia 1 μg/mL. Avibactam improved ceftaroline's MIC90s for Bacteroides fragilis from >32 to 2 μg/mL and for Prevotella spp. from >32 to 1 μg/mL. Ceftaroline alone demonstrates excellent in vitro activity against most of the aerobes found in moderate to severe DFI. The addition of avibactam provides an increased spectrum of activity including the beta-lactamase producing Prevotella, Bacteroides fragilis and ceftaroline resistant gram-negative enteric organisms.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Canine visceral leishmaniasis: diagnostic approaches based on polymerase chain reaction employing different biological samples.

  Arleana B P F Almeida, Valéria R F Sousa, Naiani D Gasparetto, Givago F R da Silva, Fabiano B Figueiredo, Valéria Dutra, Luciano Nakazato, Maria F Madeira
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  ABSTRACT: The accurate diagnosis of canine visceral leishmaniasis (CanL) is essential for visceral leishmaniasis control. To this end, DNA detection on different biological samples has been employed. In this study, we report the use of polymerase chain reaction (PCR) assay on samples such as buffy coat, bone marrow, intact skin and cutaneous ulcers fragments, and lymph node aspirate collected from 430 dogs to determine the suitable biological sample for use in CanL diagnosis. The PCR results were correlated with clinical status and other tests previously performed. Leishmania chagasi DNA was detected in 14.6% (n = 63) of the dogs investigated, regardless of the sample analyzed. Our results showed that symptomatic cases were easily diagnosed when compared to asymptomatic animals; however, the PCR proved to be very useful for Leishmania DNA detection, mainly in lymph node aspirate (41; 9.6%), irrespective of the clinical status of the dog. The finding that the lymph node aspirate produced high positivity rates and the fact that this specimen was obtained by noninvasive methods highlight its use in epidemiological survey by PCR for CanL diagnosis.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Systemic CA-MRSA infection following trauma during soccer match in inner Brazil: clinical and molecular characterization.

  Carlos Henrique Camargo, Maria de Lourdes Ribeiro de Souza da Cunha, Mariana Fávero Bonesso, Fabiana Picoli da Cunha, Alexandre Naime Barbosa, Carlos Magno Castelo Branco Fortaleza
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  ABSTRACT: Even though community-acquired methicillin resistant Staphylococcus aureus (CA-MRSA) was described a decade ago, reports from Brazil are scarce and cases occurred in large urban centers. We report MRSA sepsis in a 16-year-old male from a small town and who had no history of exposure to healthcare or recent travel. After trauma during a soccer match, he presented swelling in the right thigh, which evolved in a month to cellulitis complicated by local abscess, orchitis and pneumonia. The patient presented severe sepsis, with fever and respiratory failure. Laboratory findings included blood leukocyte counts above 40,000/mm(3) and thrombocytopenia. He was submitted to mechanical ventilation and therapy with vancomycin and imipenem. He had a slow but favorable response to therapy and was discharged after six weeks of hospitalization. MRSA grew from blood cultures and respiratory aspirates obtained before antimicrobial therapy. The isolate belonged to sequence type 5, spa type t311, harbored SCCmec type IV and genes for Panton-Valentine leukocidin and Enterotoxin A. The pulsed-field gel electrophoresis pattern was distinct from North American classic CA-MRSA clones. However, the sequence type and the spa type revealed that the clone belong to the same clonal complex isolated in Argentina. This is the first CA-MRSA infection reported in that region, with significant epidemiologic and clinical implications.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Cunninghamella echinulata causing fatally invasive fungal sinusitis.

  Robert E Leblanc, Zina Meriden, Deanna A Sutton, Elizabeth H Thompson, Dionissios Neofytos, Sean X Zhang
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  ABSTRACT: We report a fatal case of invasive fungal sinusitis caused by Cunninghamella echinulata in a febrile, neutropenic 15-year-old male with relapsing acute leukemia. The isolate was recovered from a nasal biopsy from the right middle meatus, and microscopic examination of the tissue revealed angioinvasion and necrosis. Human infection caused by this organism has not been well documented; however, this report alerts us to its life-threatening potential.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Evaluation of the BD MAX Cdiff assay for the detection of the toxin B gene of Clostridium difficile out of faecal specimens.

  Eva Leitner, Michaela Einetter, Andrea J Grisold, Egon Marth, Gebhard Feierl
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  ABSTRACT: In this evaluation study, 180 faecal specimens were tested in parallel with the new BD MAX™ Cdiff assay, the Premier™ Toxins A&B, and toxigenic culture as reference method. BD MAX™ Cdiff presented a high sensitivity (96.0%) and specificity (99.4%) with a negative predictive value of 99.4%.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Molecular concordance of concurrent Candida albicans candidemia and candiduria.

  Po-Yen Huang, Min-Hui Hung, Shian-Sen Shie, Lin-Hui Su, Ke-Yuan Chen, Jung-Jr Ye, Ping-Cheng Chiang, Hsieh-Shong Leu, Ching-Tai Huang
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  ABSTRACT: The significance of candiduria remains unclear. We correlated Candida albicans candidemia with candiduria by molecular genotyping. 33 pairs of concurrent blood and urine C. albicans isolates from 31 adult (≥18 years) were genotyped with infrequent-restriction-site PCR. The molecular concordance rates of three major genotypes were 100% for I, 82% for II, and 71% for III. The molecular concordance between concurrent C. albicans candidemia and candiduria was frequent. Our findings substantiate the importance of candiduria in appropriate clinical context as the majority of our patients were from intensive care units.
  Diagnostic microbiology and infectious disease 04/2013;
• Article: Fatal disseminated Scedosporium prolificans infection initiated by ophthalmic involvement in a patient with acute myeloblastic leukemia.

  Roberto Reinoso, Ester Carreño, Denise Hileeto, Alfredo Corell, J Carlos Pastor, Mónica Cabrero, Lourdes Vázquez, Margarita Calonge
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  ABSTRACT: Scedosporium prolificans is an opportunistic saprophytic fungus that rapidly disseminates and is intrinsically resistant to currently available antifungal drugs. We report a fatal case of disseminated S. prolificans infection that started with orbital and ocular involvement in a patient with secondary acute myeloblastic leukemia.
  Diagnostic microbiology and infectious disease 04/2013;