Development of a One-Step Multiplex PCR Assay for Differential Detection of Major Mycobacterium Species.

Abstract:

:The prevalence of tuberculosis continues to be high, and nontuberculous mycobacterial (NTM) infection has also emerged worldwide. Moreover, differential and accurate identification of mycobacteria to the species or subspecies level is an unmet clinical need. Here, we developed a one-step multiplex PCR assay using whole-genome analysis and bioinformatics to identify novel molecular targets. The aims of this assay were to (i) discriminate between the Mycobacterium tuberculosis complex (MTBC) and NTM using rv0577 or RD750, (ii) differentiate M. tuberculosis (M. tuberculosis) from MTBC using RD9, (iii) selectively identify the widespread M. tuberculosis Beijing genotype by targeting mtbk_20680, and (iv) simultaneously detect five clinically important NTM (M. avium, M. intracellulare, M. abscessus, M. massiliense, and M. kansasii) by targeting IS1311, DT1, mass_3210, and mkan_rs12360 An initial evaluation of the multiplex PCR assay using reference strains demonstrated 100% specificity for the targeted Mycobacterium species. Analytical sensitivity ranged from 1 to 10 pg for extracted DNA and was 103 and 104 CFU for pure cultures and nonhomogenized artificial sputum cultures, respectively, of the targeted species. The accuracy of the multiplex PCR assay was further evaluated using 55 reference strains and 94 mycobacterial clinical isolates. Spoligotyping, multilocus sequence analysis, and a commercial real-time PCR assay were employed as standard assays to evaluate the multiplex PCR assay with clinical M. tuberculosis and NTM isolates. The PCR assay displayed 100% identification agreement with the standard assays. Our multiplex PCR assay is a simple, convenient, and reliable technique for differential identification of MTBC, M. tuberculosis, M. tuberculosis Beijing genotype, and major NTM species.

journal_name

J Clin Microbiol

authors

Chae H,Han SJ,Kim SY,Ki CS,Huh HJ,Yong D,Koh WJ,Shin SJ

doi

10.1128/JCM.00549-17

subject

Has Abstract

pub_date

2017-09-01 00:00:00

pages

2736-2751

issue

9

eissn

0095-1137

issn

1098-660X

pii

JCM.00549-17

journal_volume

55

pub_type

杂志文章
  • Cas12a/Guide RNA-Based Platform for Rapid and Accurate Identification of Major Mycobacterium Species.

    abstract::Mycobacterium tuberculosis infection and nontuberculous mycobacteria (NTM) infections exhibit similar clinical symptoms; however, the therapies for these two types of infections are different. Therefore, the rapid and accurate identification of M. tuberculosis and NTM species is very important for the control of tuber...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01368-19

    authors: Xiao G,He X,Zhang S,Liu Y,Liang Z,Liu H,Zhang J,Ou M,Cai S,Lai W,Zhang T,Ren L,Zhang G

    更新日期:2020-01-28 00:00:00

  • Detection by polymerase chain reaction amplification of human herpesvirus 6 DNA in peripheral blood of patients with exanthem subitum.

    abstract::The polymerase chain reaction was used to detect human herpesvirus 6 (HHV-6) DNA in peripheral blood mononuclear cells from patients with exanthem subitum. Amplified products were detected by agarose gel electrophoresis and dot blot hybridization with a cloned DNA probe. No cross-hybridization with DNAs of five other ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.5.970-974.1990

    authors: Kondo K,Hayakawa Y,Mori H,Sato S,Kondo T,Takahashi K,Minamishima Y,Takahashi M,Yamanishi K

    更新日期:1990-05-01 00:00:00

  • Rapid detection of Helicobacter pylori in gastric biopsy material by polymerase chain reaction.

    abstract::By using primers based on the sequence of a species-specific antigen of Helicobacter pylori (P. O'Toole, S.M. Logan, M. Kostrzynska. T. Wadström, and T.J. Trust, J. Bacteriol. 173:505-513, 1991), a protocol was established for detection of this microorganism in gastric biopsy samples by the polymerase chain reaction (...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.30.1.54-58.1992

    authors: Hammar M,Tyszkiewicz T,Wadström T,O'Toole PW

    更新日期:1992-01-01 00:00:00

  • Relationship between mutations in dihydropteroate synthase of Pneumocystis carinii f. sp. hominis isolates in Japan and resistance to sulfonamide therapy.

    abstract::We examined mutations in the dihydropteroate synthase (DHPS) genes of Pneumocystis carinii f. sp. hominis (P. carinii) strains isolated from 24 patients with P. carinii pneumonia (PCP) in Japan. DHPS mutations were identified at amino acid positions 55 and/or 57 in isolates from 6 (25.0%) of 24 patients. The underlyin...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.38.9.3161-3164.2000

    authors: Takahashi T,Hosoya N,Endo T,Nakamura T,Sakashita H,Kimura K,Ohnishi K,Nakamura Y,Iwamoto A

    更新日期:2000-09-01 00:00:00

  • Infectious but nonpathogenic isolate of Borrelia burgdorferi.

    abstract::We document for the first time an infectious but nonarthritogenic variant of Borrelia burgdorferi. Strain 25015, previously isolated from an Ixodes dammini larva collected in upstate New York, was infectious but failed to produce arthritis or carditis in laboratory rats and mice. By contrast, pathogenic strain N40 inv...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.12.2693-2699.1990

    authors: Anderson JF,Barthold SW,Magnarelli LA

    更新日期:1990-12-01 00:00:00

  • Mechanism of cross-contamination of blood culture bottles in outbreaks of pseudobacteremia associated with nonsterile blood collection tubes.

    abstract::Several studies have identified nonsterile blood collection tubes as the reservoir for contaminating microorganisms in outbreaks of pseudobacteremia. Microorganisms are thought to be transferred from the nonsterile tubes to the blood culture tubes when blood is drawn for cultures, but the validity of this hypothesis h...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.22.1.23-25.1985

    authors: McNeil MM,Davis BJ,Anderson RL,Martone WJ,Solomon SL

    更新日期:1985-07-01 00:00:00

  • Isolation of influenza virus in human lung embryonated fibroblast cells (MRC-5) from clinical samples.

    abstract::Ninety-four pharyngeal swab samples corresponding to 94 patients with suspected influenza virus infection were inoculated in Madin-Darby canine kidney (MDCK) cells, the conventional cell system for the isolation of influenza virus, and in fibroblastic human embryo lung (MRC-5) cells, a cell system less commonly used f...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.7.1948-1949.1995

    authors: de Oña M,Melón S,de la Iglesia P,Hidalgo F,Verdugo AF

    更新日期:1995-07-01 00:00:00

  • Salivary immunoglobulin G assay to diagnose Helicobacter pylori infection in children.

    abstract::An in-house enzyme-linked immunosorbent assay (ELISA) for measurement of Helicobacter pylori-specific immunoglobulin G (IgG) and IgA in saliva was evaluated by comparison with histopathologic (Giemsa staining) and biochemical (urease quick test) examination of gastric biopsy specimens obtained from 112 children referr...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.35.12.3358-3360.1997

    authors: Luzza F,Oderda G,Maletta M,Imeneo M,Mesuraca L,Chioboli E,Lerro P,Guandalini S,Pallone F

    更新日期:1997-12-01 00:00:00

  • Molecular typing of Aspergillus terreus isolates collected in Houston, Texas, and Innsbruck, Austria: evidence of great genetic diversity.

    abstract::Aspergillus terreus isolates collected from patients at The M. D. Anderson Cancer Center in Houston, TX, and at The University Hospital of Innsbruck, Austria, were analyzed using random amplification of polymorphic DNA-PCR with three different primers. No strain similarity in either institution was detected, indicatin...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00917-07

    authors: Lass-Flörl C,Grif K,Kontoyiannis DP

    更新日期:2007-08-01 00:00:00

  • Evaluation of a novel dry latex preparation for demonstration of infectious mononucleosis heterophile antibody in comparison with three established tests.

    abstract::A new, dried antigen-coated latex preparation for the demonstration of infectious mononucleosis (IM) heterophile antibody (Dryspot IM kit; Oxoid, Ltd., Basingstoke, Hampshire, United Kingdom) was compared with the IM kit (a liquid latex reagent from the same source), an immunoassay (ImmunoCard Mono; Meridian Diagnosti...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.37.1.95-98.1999

    authors: Rogers R,Windust A,Gregory J

    更新日期:1999-01-01 00:00:00

  • Variable serum immunoglobulin responses against different Borrelia burgdorferi sensu lato species in a population at risk for and patients with Lyme disease.

    abstract::Borrelia burgdorferi sensu lato species display considerable antigenic polymorphism. In order to evaluate the importance of this antigenic heterogeneity in the serodiagnosis of Lyme disease, the serum immunoglobulin G response in 148 healthy individuals from an area in northern Sweden where Lyme disease is endemic and...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.6.1473-1478.1995

    authors: Bunikis J,Olsén B,Westman G,Bergstroöm S

    更新日期:1995-06-01 00:00:00

  • Direct Detection of Rifampin and Isoniazid Resistance in Sputum Samples from Tuberculosis Patients by High-Resolution Melt Curve Analysis.

    abstract::Drug-resistant tuberculosis (TB) is a major threat to TB control worldwide. Globally, only 40% of the 340,000 notified TB patients estimated to have multidrug-resistant-TB (MDR-TB) were detected in 2015. This study was carried out to evaluate the utility of high-resolution melt curve analysis (HRM) for the rapid and d...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02104-16

    authors: Anthwal D,Gupta RK,Bhalla M,Bhatnagar S,Tyagi JS,Haldar S

    更新日期:2017-06-01 00:00:00

  • Aerobically incubated medium for decarboxylase testing of Enterobacteriaceae by replica-plating methods.

    abstract::An aerobically incubated, agar-based medium was developed for amino acid decarboxylase testing of Enterobacteriaceae family members by replica-plating methods. Results with the new medium agreed 97 to 99% with the reference broth method of Moeller, and no false-positive reactions were encountered. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.10.6.940-942.1979

    authors: Maccani JE

    更新日期:1979-12-01 00:00:00

  • Evaluation of a multiplex real-time reverse transcriptase PCR assay for detection and differentiation of influenza viruses A and B during the 2001-2002 influenza season in Israel.

    abstract::The ability to rapidly diagnose influenza virus infections is of the utmost importance in the evaluation of patients with upper respiratory tract infections. It is also important for the influenza surveillance activities performed by national influenza centers. In the present study we modified a multiplex real-time re...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.43.2.589-595.2005

    authors: Hindiyeh M,Levy V,Azar R,Varsano N,Regev L,Shalev Y,Grossman Z,Mendelson E

    更新日期:2005-02-01 00:00:00

  • Latex agglutination test for monitoring antibodies to avian influenza virus subtype H5N1.

    abstract::A latex agglutination test (LAT) based on polystyrene beads sensitized with inactivated avian influenza virus H5N1 particles was developed. Compared with the hemagglutination inhibition test, the sensitivity and specificity of the LAT were 88.8 and 97.6%, respectively, in detecting 830 serum samples from vaccinated ch...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.43.4.1953-1955.2005

    authors: Xu X,Jin M,Yu Z,Li H,Qiu D,Tan Y,Chen H

    更新日期:2005-04-01 00:00:00

  • Native valve endocarditis caused by an organism resembling Corynebacterium striatum.

    abstract::An organism resembling Corynebacterium striatum was isolated from the blood of a patient with acute aortic valvular insufficiency and no history of valvular heart disease. At autopsy, histopathologic examination of the aortic valve revealed pleomorphic gram-positive bacilli and destruction of valvular tissue. Our isol...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.1.8-10.1990

    authors: Markowitz SM,Coudron PE

    更新日期:1990-01-01 00:00:00

  • Evaluation of Ebola virus inactivation procedures for Plasmodium falciparum malaria diagnostics.

    abstract::Plasmodium falciparum malaria is highly endemic in the three most affected countries in the current epidemic of Ebola virus disease (EVD) in West Africa. As EVD and malaria are clinically indistinguishable, both remain part of the differential diagnosis of ill travelers from returning from areas of EVD transmission. W...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00165-15

    authors: Lau R,Wang A,Chong-Kit A,Ralevski F,Boggild AK

    更新日期:2015-04-01 00:00:00

  • Chlamydia trachomatis serovar distribution and Neisseria gonorrhoeae coinfection in male patients with urethritis in Greece.

    abstract::The distribution of Chlamydia trachomatis serovars and Neisseria gonorrhoeae coinfection was studied in a group of 100 C. trachomatis-positive males with urethritis in Greece. The serovar distribution revealed that apart from the predominant worldwide types E and F, the relatively uncommon type G is also prevalent. Go...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00586-10

    authors: Papadogeorgakis H,Pittaras TE,Papaparaskevas J,Pitiriga V,Katsambas A,Tsakris A

    更新日期:2010-06-01 00:00:00

  • Comparison of PCR-ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for typing Clostridium difficile.

    abstract::Clostridium difficile is now recognized as the major agent responsible for nosocomial diarrhea in adults. Among the genotyping methods available, arbitrarily primed PCR (AP-PCR), PCR-ribotyping, and pulsed-field gel electrophoresis (PFGE) have been widely used for investigating outbreaks of C. difficile infections. Ho...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.38.7.2484-2487.2000

    authors: Bidet P,Lalande V,Salauze B,Burghoffer B,Avesani V,Delmée M,Rossier A,Barbut F,Petit JC

    更新日期:2000-07-01 00:00:00

  • Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

    abstract::The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, inc...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.27.7.1623-1625.1989

    authors: Walpita P,Darougar S

    更新日期:1989-07-01 00:00:00

  • Multiplex PCR allows rapid and accurate diagnosis of bloodstream infections in newborns and children with suspected sepsis.

    abstract::Sepsis is a major health problem in newborns and children. Early detection of pathogens allows initiation of appropriate antimicrobial therapy that strongly correlates with positive outcomes. Multiplex PCR has the potential to rapidly identify bloodstream infections, compensating for the loss of blood culture sensitiv...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02460-10

    authors: Lucignano B,Ranno S,Liesenfeld O,Pizzorno B,Putignani L,Bernaschi P,Menichella D

    更新日期:2011-06-01 00:00:00

  • Colonization of human wounds by Escherichia vulneris and Escherichia hermannii.

    abstract::In this report we present clinical descriptions of 12 Hawaiian patients from whom Escherichia vulneris or E. hermannii strains were isolated. All but two patients had soft-tissue infections with multiple bacteria, particularly Staphylococcus aureus. The other two had purulent conjunctivitis associated with S. aureus a...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.22.2.283-285.1985

    authors: Pien FD,Shrum S,Swenson JM,Hill BC,Thornsberry C,Farmer JJ 3rd

    更新日期:1985-08-01 00:00:00

  • Assessment of enzyme immunoassay and immunofluorescence tests for detection of Chlamydia trachomatis.

    abstract::Two rapid diagnostic tests for Chlamydia trachomatis (Microtrak, Syva Co., Palo Alto, Calif.; and Chlamydiazyme, Abbott Laboratories, North Chicago, Ill.) were evaluated in comparison with growth of the organism in tissue culture for 2,030 urogenital specimens from men and women. Neither test performed as well as cult...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.25.10.1938-1943.1987

    authors: Hipp SS,Han Y,Murphy D

    更新日期:1987-10-01 00:00:00

  • Comparison of the BD MAX enteric bacterial panel to routine culture methods for detection of Campylobacter, enterohemorrhagic Escherichia coli (O157), Salmonella, and Shigella isolates in preserved stool specimens.

    abstract::We compared the BD MAX enteric bacterial panel (EBP) to culture for the detection of Salmonella, Shigella, Campylobacter, and Shiga toxin-producing enterohemorrhagic Escherichia coli (EHEC) O157 in seeded stool samples. The EBP panel demonstrated superior sensitivity and reliably detected Salmonella, EHEC O157, Shigel...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.03099-13

    authors: Anderson NW,Buchan BW,Ledeboer NA

    更新日期:2014-04-01 00:00:00

  • Performance of Antigen Concentration Thresholds for Attributing Fever to Malaria among Outpatients in Angola.

    abstract::The density of malaria parasites is a key determinant of whether an infected individual develops fever. While the pyrogenic threshold for malaria parasite density has been well studied, there are no analogous data on the antigen levels associated with fever during infection. Samples from 797 afebrile and 457 febrile o...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01901-18

    authors: Plucinski MM,Rogier E,Dimbu PR,Fortes F,Halsey ES,Aidoo M,Smith T

    更新日期:2019-02-27 00:00:00

  • Inverse association between Lancefield group G Streptococcus colonization and sore throat in slum and nonslum settings in Brazil.

    abstract::Group G Streptococcus has been implicated as a causative agent of pharyngitis in outbreak situations, but its role in endemic disease remains elusive. We found an unexpected inverse association of Streptococcus dysgalactiae subsp. equisimilis colonization and sore throat in a study of 2,194 children of 3 to 15 years o...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02095-10

    authors: Tartof SY,Farrimond F,de Matos JA,Reis JN,Ramos RT,Andrade AN,dos Reis MG,Riley LW

    更新日期:2011-01-01 00:00:00

  • Comparison of ortho respiratory syncytial virus enzyme-linked immunosorbent assay and HEp-2 cell culture.

    abstract::Two hundred seventy nasopharyngeal aspirates were tested in duplicate with the Ortho Diagnostics, Inc. (Raritan, N.J.), respiratory syncytial virus antigen enzyme-linked immunosorbent assay. The test was sensitive (80 to 82%) and specific (96%) when compared with cell culture. The enzyme-linked immunosorbent assay det...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.22.6.1071-1072.1985

    authors: Bromberg K,Tannis G,Daidone B,Clarke L,Sierra MF

    更新日期:1985-12-01 00:00:00

  • Impact of long-term storage on stability of standard DNA for nucleic acid-based methods.

    abstract::Real-time PCR is dependent upon a calibration function for quantification. While long-term storage of standards saves cost and time, solutions of DNA are prone to degradation. We present here the benchmark treatment for preservation of DNA standards, involving storage in 50% glycerol-double-distilled water, whereby a ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01230-10

    authors: Röder B,Frühwirth K,Vogl C,Wagner M,Rossmanith P

    更新日期:2010-11-01 00:00:00

  • Molecular analysis of microflora associated with dentoalveolar abscesses.

    abstract::The microflora associated with three dentoalveolar abscesses was determined by cultural and molecular methods. 16S rRNA genes were randomly amplified by means of conserved eubacterial primers and cloned. Restriction fragment length polymorphism analysis of the clones and amplified genes encoding 16S rRNA from the cult...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.34.3.537-542.1996

    authors: Dymock D,Weightman AJ,Scully C,Wade WG

    更新日期:1996-03-01 00:00:00

  • Mannheimia species associated with ovine mastitis.

    abstract::Mannheimia glucosida, M. haemolytica, and M. ruminalis were isolated from cases of acute mastitis in ewes. M. glucosida was found to be a common cause of clinical mastitis in sheep. Selected phenotypic tests in addition to genotyping were needed to definitively identify Mannheimia species causing ovine mastitis. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01145-10

    authors: Omaleki L,Barber SR,Allen JL,Browning GF

    更新日期:2010-09-01 00:00:00