Direct detection of Mycobacterium tuberculosis in clinical specimens using single-tube biotinylated nested polymerase chain reaction-enzyme linked immunoassay (PCR-ELISA).


:A biotinylated single-tube nested polymerase chain reaction (PCR) assay with microwell hybridization assay (bPCR-ELISA) was developed for detection of Mycobacterium tuberculosis in clinical specimens. A total of 659 specimens (601 respiratory specimens and 58 nonrespiratory specimens) were collected for evaluation using three DNA amplification techniques: newly designed bPCR-ELISA, in-house single-tube nested PCR for IS6110 gene sequence (nPCR), and commercial automated assays, the Cobas Amplicor System from Roche Diagnostic Systems (aPCR). Sixty-four (9.7%) specimens were culture-positive for M. tuberculosis. Eleven (1.7%) specimens culture-positive for nontuberculosis mycobacteria were negative by all three PCR assays. The resolved performance of bPCR-ELISA, nPCR, and aPCR was found at sensitivities of 97%, 94%, and 97%, respectively. All three PCR assays exhibited a 100% specificity. In evaluation of bPCR-ELISA, a clear distinction between PCR-positive and PCR-negative specimens when an OD405 value of 0.6 was chosen as cut-off. With serial dilutions of M. tuberculosis H37Rv DNA, the detection limit of bPCR-ELISA was found to be 0.75 cfu per reaction at OD405 value of 0.6. Our developed bPCR-ELISA provides a highly sensitive and low-costing molecular diagnosis suitable for developing countries with high prevalence of tuberculosis.


Yam WC,Cheng VC,Hui WT,Wang LN,Seto WH,Yuen KY




Has Abstract


2004-04-01 00:00:00














  • Detection of Pneumocystis carinii by DNA amplification in human immunodeficiency virus-positive patients.

    abstract::The opportunistic pathogen Pneumocystis carinii (PC) is a frequent cause of a life-threatening pneumonia in human immunodeficiency virus (HIV)-infected individuals and in other immunocompromised hosts. Specimens obtained from 128 bronchoalveolar lavage (BAL) fluid samples from 123 HIV-positive patients with pulmonary ...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 临床试验,杂志文章


    authors: Savoia D,Millesimo M,Cassetta I,Forno B,Caramello P

    更新日期:1997-10-01 00:00:00

  • Multicenter evaluation of the Verigene Gram-negative blood culture nucleic acid test for rapid detection of bacteria and resistance determinants in positive blood cultures.

    abstract::The Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN) is a microarray-based assay that enables rapid detection of 9 common Gram-negative bacteria and 6 resistance determinants directly from positive blood cultures. We compared the performance of BC-GN with currently used automated systems, testing 141 cli...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章,多中心研究


    authors: Uno N,Suzuki H,Yamakawa H,Yamada M,Yaguchi Y,Notake S,Tamai K,Yanagisawa H,Misawa S,Yanagihara K

    更新日期:2015-12-01 00:00:00

  • Development of a diagnostic real-time polymerase chain reaction assay for the detection of invasive Haemophilus influenzae in clinical samples.

    abstract::Since the introduction of the Haemophilus influenzae serotype b vaccine, invasive H. influenzae disease has become dominated by nontypeable (NT) strains. Several widely used molecular diagnostic methods have been shown to lack sensitivity or specificity in the detection of some of these strains. Novel real-time assays...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Meyler KL,Meehan M,Bennett D,Cunney R,Cafferkey M

    更新日期:2012-12-01 00:00:00

  • Proteolytic activity of Candida albicans and other yeasts.

    abstract::Clinical isolates of C. albicans (75 strains) and other yeasts (20 strains) were evaluated for their ability to produce a carboxyl acid proteinase in an effort to assess its potential role as a virulence factor. Yeasts were categorized as to the infectious process present in the patient: (1) isolates from patients wit...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Schreiber B,Lyman CA,Gurevich J,Needham CA

    更新日期:1985-01-01 00:00:00

  • Epidemiology and antibiotic susceptibility of aerococci in urinary cultures.

    abstract::In this study, we present population-based data regarding the prevalence of aerococci in clinical urinary samples. During a 3-month period, all aerococcal isolates from urinary samples from 2 clinical microbiology laboratories were collected. We identified 64 Aerococcus urinae isolates and 40 Aerococcus sanguinicola i...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Senneby E,Petersson AC,Rasmussen M

    更新日期:2015-02-01 00:00:00

  • Comparison of community-associated and health care-associated methicillin-resistant Staphylococcus aureus in Canada: results of the CANWARD 2007-2009 study.

    abstract::This study assessed the demographics, antimicrobial susceptibility, and molecular epidemiology of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) and health care-associated MRSA (HA-MRSA) in Canadian hospitals between 2007 and 2009. Among 3589 S. aureus, 889 (24.8%) were MRSA; 224 (25.2%) we...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Nichol KA,Adam HJ,Hussain Z,Mulvey MR,McCracken M,Mataseje LF,Thompson K,Kost S,Lagacé-Wiens PR,Hoban DJ,Zhanel GG,Canadian Antimicrobial Resistance Alliance (CARA).

    更新日期:2011-03-01 00:00:00

  • Bipolaris oryzae, a novel fungal opportunist causing keratitis.

    abstract::We report a case of mycotic keratitis caused by Bipolaris oryzae with predisposing trauma from a foreign body. The fungus was identified by sequencing the internal transcribed spacer region, translation elongation factor 1α (TEF1) gene, and partial glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene, and the species ...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Wang L,Al-Hatmi AM,Lai X,Peng L,Yang C,Lai H,Li J,Meis JF,de Hoog GS,Zhuo C,Chen M

    更新日期:2016-05-01 00:00:00

  • Partial comparison of the NxTAG Respiratory Pathogen Panel Assay with the Luminex xTAG Respiratory Panel Fast Assay V2 and singleplex real-time polymerase chain reaction for detection of respiratory pathogens.

    abstract::In this study, 185 nasopharyngeal swabs were tested to compare the sensitivity and specificity of the Luminex NxTAG (NxTAG) Respiratory Pathogen Panel (RPP) Assay with those of the Luminex Respiratory Virus Panel (RVP) Fast Assay v2 and singleplex real-time polymerase chain reaction (PCR). The NxTAG Assay identified a...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Esposito S,Scala A,Bianchini S,Presicce ML,Mori A,Sciarrabba CS,Fior G,Principi N

    更新日期:2016-09-01 00:00:00

  • Antimicrobial susceptibility and extended-spectrum beta-lactamase rates in aerobic gram-negative bacteria causing intra-abdominal infections in Vietnam: report from the Study for Monitoring Antimicrobial Resistance Trends (SMART 2009-2011).

    abstract::Treatment options for multidrug-resistant pathogens remain problematic in many regions and individual countries, warranting ongoing surveillance and analysis. Limited antimicrobial susceptibility information is available for pathogens from Vietnam. This study determined the bacterial susceptibility of aerobic gram-neg...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Biedenbach DJ,Bouchillon SK,Hoban DJ,Hackel M,Phuong DM,Nga TT,Phuong NT,Phuong TT,Badal RE

    更新日期:2014-08-01 00:00:00

  • Evaluation of the Copan ESwab transport system for the detection of methicillin-resistant Staphylococcus aureus: a laboratory and clinical study.

    abstract::Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as an important pathogen. Successful detection starts with the selection of an appropriate specimen collection device. The Copan ESwab, a new swab, was compared with a commonly used Copan Venturi swab for the recovery of MRSA. In vitro assessment was perfo...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Smismans A,Verhaegen J,Schuermans A,Frans J

    更新日期:2009-10-01 00:00:00

  • SPM-1-producing Pseudomonas aeruginosa ST277 clone recovered from microbiota of migratory birds.

    abstract::The production of São Paulo metallo-β-lactamase (SPM-1) is the most common carbapenem resistance mechanism detected among multidrug-resistant Pseudomonas aeruginosa clinical isolates in Brazil. Dissemination of SPM-1-producing P. aeruginosa has been restricted to the nosocomial settings, with sporadic reports of envir...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Martins WMBS,Narciso AC,Cayô R,Santos SV,Fehlberg LCC,Ramos PL,da Cruz JB,Gales AC

    更新日期:2018-03-01 00:00:00

  • A novel OXA-10-like beta-lactamase is present in different Enterobacteriaceae.

    abstract::OXA 101, a novel OXA-10 like enzyme, was found forming part of a class 1 integron located in a conjugative plasmid in three different species of Enterobacteriaceae. This beta-lactamase is related to OXA-35 and OXA-56 and displays a narrow substrate hydrolysis profile. ...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Porto A,Ayala J,Gutkind G,Di Conza J

    更新日期:2010-02-01 00:00:00

  • The use of biotyping and DNA fingerprinting in typing Candida albicans from hospitalized patients.

    abstract::The application of typing procedures for the purpose of strain differentiation among isolates of Candida albicans obtained from hospitalized patients has been limited. We have applied biotyping and DNA restriction fragment analysis (DNA fingerprinting) by using EcoRI to the study of C. albicans isolates obtained from ...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Pfaller MA,Cabezudo I,Hollis R,Huston B,Wenzel RP

    更新日期:1990-11-01 00:00:00

  • Igm recognition of recombinant Toxoplasma gondii antigens by sera of acutely or latently infected humans.

    abstract::Clinical non-relevant (CNR) IgM specific for Toxoplasma gondii is responsible for false-positive results in commercially available IgM assays. Using IgM immunoblotting, it is possible to distinguish between IgM in sera of acutely infected (AI) patients and CNR IgM. Especially the combination of staining of a 55 and 30...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Meek B,Diepersloot RJ,van Gool T,Speijer D,Peek R

    更新日期:2003-01-01 00:00:00

  • Evaluation of molecular methods for detection of Borrelia burgdorferi senso lato in ticks.

    abstract::Borrelia burgdorferi sensu lato (s. l.), the agent of Lyme disease, is distributed widely worldwide. A large number of polymerase chain reaction (PCR) methods have been developed and used for detection of B. burgdorferi s. l. However, there is a lack of a reference standard because of the genetic diversity of the B. b...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Yang J,Liu Z,Guan G,Che R,Niu Q,Li Y,Liu J,Ma M,Ren Q,Liu A,Luo J,Yin H

    更新日期:2012-05-01 00:00:00

  • Direct detection of Streptococcus pneumoniae in positive blood cultures by real-time polymerase chain reaction.

    abstract::We developed a real-time polymerase chain reaction specific for Streptococcus pneumoniae to be applied directly from blood culture bottles without previous DNA extraction step. For the 128 blood culture bottles tested, the assay had 94% and 98.4% sensitivity and specificity, respectively. This assay provides rapid and...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Selva L,Esteva C,Gené A,de Sevilla MF,Hernandez-Bou S,Muñoz-Almagro C

    更新日期:2010-02-01 00:00:00

  • In vitro activities of voriconazole, posaconazole, and fluconazole against 4,169 clinical isolates of Candida spp. and Cryptococcus neoformans collected during 2001 and 2002 in the ARTEMIS global antifungal surveillance program.

    abstract::We examined the in vitro activities of voriconazole, posaconazole, and fluconazole against 3,932 isolates of Candida spp. and 237 isolates of Cryptococcus neoformans obtained from over 100 medical centers worldwide during 2001 and 2002. The MICs of the antifungal drugs were determined by broth microdilution tests perf...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Pfaller MA,Messer SA,Boyken L,Hollis RJ,Rice C,Tendolkar S,Diekema DJ

    更新日期:2004-03-01 00:00:00

  • Impact of AmpC overexpression on outcomes of patients with Pseudomonas aeruginosa bacteremia.

    abstract::AmpC overexpression (AmpC++) is a significant mechanism of beta-lactam resistance in Pseudomonas aeruginosa, but its impact on clinical outcomes is not well established. To examine the influence of AmpC++ on clinical outcomes of patients with P. aeruginosa bacteremia, we screened all bloodstream P. aeruginosa isolates...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Tam VH,Chang KT,Schilling AN,LaRocco MT,Genty LO,Garey KW

    更新日期:2009-03-01 00:00:00

  • Four year prospective evaluation of nosocomial bacteremia: epidemiology, microbiology, and patient outcome.

    abstract::A prospective study of all patients with clinically significant nosocomial bacteremia at one institution from 1994 to 1997 was performed to: (1) describe the epidemiology and microbiology of nosocomial bacteremias; (2) determine the crude mortality associated with such infections; and (3) identify independent predicto...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Lark RL,Chenoweth C,Saint S,Zemencuk JK,Lipsky BA,Plorde JJ

    更新日期:2000-11-01 00:00:00

  • Varicella associated intracerebral hemorrhage in the absence of thrombocytopenia.

    abstract::A young woman with varicella complicated by streptococcal cellulitis developed an intracerebral bleed in the absence of thrombocytopenia, serious coagulopathy, or a detectable vascular lesion. We postulate that during the acute infection with varicella, a cerebral endothelial lesion was produced which later caused the...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Fikrig E,Barg NL

    更新日期:1989-07-01 00:00:00

  • Methicillin-susceptible strains responsible for postoperative orthopedic infection are not selected by the use of cefazolin in prophylaxis.

    abstract::Comparison of methicillin-susceptible Staphylococcus aureus (MSSA) isolates responsible for bone and joint infection (BJI, n=73) and nasal colonization (n=57) revealed similar prevalence of β-lactamase (blaZ) type A production, associated with cefazolin hydrolysis, suggesting that blaZ type A-carrying MSSA isolates im...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Trouillet-Assant S,Valour F,Mouton W,Martins-Simões P,Lustig S,Laurent F,Ferry T,Lyon BJI study group.

    更新日期:2016-03-01 00:00:00

  • Polymicrobial candidemia.

    abstract::We conducted a retrospective review of the clinical data on patients with polymicrobial candidemia during a nine-year period (1992-2000) at our tertiary care University Hospital. Also, the clinical features of patients with polymicrobial vs monomicrobial candidemia were compared. There were 16 (5.2%) patients with mul...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章,评审


    authors: Pulimood S,Ganesan L,Alangaden G,Chandrasekar P

    更新日期:2002-12-01 00:00:00

  • Use of cerebrospinal fluid shunt for the management of elevated intracranial pressure in a patient with active AIDS-related cryptococcal meningitis.

    abstract::Persistently elevated intracranial pressure (ICP) is one of the most accurate predictors of a poor prognosis in patients with AIDS-related cryptococcal meningitis. We present a severe case of persistent cryptococcal meningitis in a patient with advanced AIDS, complicated by elevation of ICP. A ventriculoperitoneal shu...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Mylonakis E,Merriman NA,Rich JD,Flanigan TP,Walters BC,Tashima KT,Mileno MD,van der Horst CM

    更新日期:1999-06-01 00:00:00

  • In vitro activity of fosfomycin against ciprofloxacin-resistant or extended-spectrum beta-lactamase-producing Escherichia coli isolated from urine and blood.

    abstract::In this study, we evaluated the in vitro activity of fosfomycin and 7 other comparator agents against 307 Escherichia coli isolates including ciprofloxacin-resistant or extended-spectrum beta-lactamase (ESBL)-producing isolates. Bacterial isolates were collected from urine and blood from patients at a Korean tertiary-...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Ko KS,Suh JY,Peck KR,Lee MY,Oh WS,Kwon KT,Jung DS,Lee NY,Song JH

    更新日期:2007-05-01 00:00:00

  • In vitro activity of the novel ketolide HMR 3647 and comparative oral antibiotics against Canadian respiratory tract isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis.

    abstract::The in vitro activities of HMR 3647, erythromycin A, clarithromycin, azithromycin, roxithromycin, penicillin G, ampicillin, cefuroxime, trimethoprim/sulfamethoxazole, tetracycline, ciprofloxacin, and levofloxacin were determined for 1179 Streptococcus pneumoniae, 1438 Haemophilus influenzae, and 428 Moraxella catarrha...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章,多中心研究


    authors: Hoban DJ,Zhanel GG,Karlowsky JA

    更新日期:1999-09-01 00:00:00

  • Activity of ceftazidime-avibactam against Escherichia coli isolates from U.S. veterans (2011) in relation to co-resistance and sequence type 131 (ST131) H30 and H30Rx status.

    abstract::Escherichia coli ST131, with its multidrug-resistance-associated H30R1 and H30Rx clonal subsets within the H30R subclone, causes most antimicrobial-resistant E. coli infections. The activity of ceftazidime-avibactam (CZA) against ST131 strains is undefined. We determined CZA MICs for 595 E. coli clinical isolates from...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Johnston BD,Thuras PD,Johnson JR

    更新日期:2020-07-01 00:00:00

  • Susceptibility testing of Mycobacterium tuberculosis by broth microdilution method: a rapid alternative method.

    abstract::A total of 50 clinical isolates of Mycobacterium tuberculosis were tested by the proportion method and the broth microdilution method (BMM) for primary antitubercular drugs. The results were obtained after 14 days of incubation. There was a 100% agreement for all drugs except streptomycin, which showed a 96% agreement...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Sethi S,Mandal J,Kumar P,Kumar S,Meharwal S,Jindal SK,Sharma M

    更新日期:2007-04-01 00:00:00

  • Mycobacterium avium: a pathogen of patients with acquired immunodeficiency syndrome.

    abstract::Mycobacterium avium complex has been isolated with increasing frequency from humans during the last few decades. Thirteen patients admitted to the UCLA Medical Center with the diagnosis of acquired immunodeficiency syndrome (AIDS), in addition to having Kaposi's sarcoma, Pneumocystis pneumonia, and other opportunistic...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Berlin OG,Zakowski P,Bruckner DA,Clancy MN,Johnson BL Jr

    更新日期:1984-06-01 00:00:00

  • Disseminated "Haemophilus quentini" infection in a patient with multiple myeloma - a case report and review of the literature.

    abstract::We describe a case report of a 56-year-old male with undiagnosed multiple myeloma who had severe sepsis associated with pneumonia, meningitis, polyarthritis, and osteomyelitis related to invasive "Haemophilus quentini" infection. The genus was misidentified as H. influenzae by the common bacterial identification syste...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章,评审


    authors: Cohen R,Finn T,Babushkin F,Karalnik S,Paikin S,Adler A,Geffen Y,Rokney A,Ron M

    更新日期:2019-07-01 00:00:00

  • Carry-over of host nutrients during sampling enhances undesired growth of Staphylococcus aureus in liquid Amies transport medium.

    abstract::Optimal transportation of bacteria is important for accurate clinical interpretation, quantitative assays, and microbiome studies. A transport medium should ideally keep the bacteria alive without supporting growth or altering the relative proportions of the constituent species. We investigated the effect of nasal muc...

    journal_title:Diagnostic microbiology and infectious disease

    pub_type: 杂志文章


    authors: Islam MZ,Larsen J,Skov R,Angen Ø

    更新日期:2019-01-01 00:00:00