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 Microbioljournal_title
Journal of clinical microbiologyauthors
Chae H,Han SJ,Kim SY,Ki CS,Huh HJ,Yong D,Koh WJ,Shin SJdoi
10.1128/JCM.00549-17subject
Has Abstractpub_date
2017-09-01 00:00:00pages
2736-2751issue
9eissn
0095-1137issn
1098-660Xpii
JCM.00549-17journal_volume
55pub_type
杂志文章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
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doi:10.1128/JCM.01368-19
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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
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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
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doi:10.1128/JCM.30.1.54-58.1992
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.12.2693-2699.1990
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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
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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
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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
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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
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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
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.6.1473-1478.1995
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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
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doi:10.1128/JCM.02104-16
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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
更新日期:1979-12-01 00:00:00
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
更新日期:2005-02-01 00:00:00
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
更新日期:2005-04-01 00:00:00
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
更新日期:1990-01-01 00:00:00
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
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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
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.27.7.1623-1625.1989
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.22.2.283-285.1985
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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
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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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
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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
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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
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