Abstract:
:Current nucleic acid amplification methods to detect Mycobacterium tuberculosis are complex, labor-intensive, and technically challenging. We developed and performed the first analysis of the Cepheid Gene Xpert System's MTB/RIF assay, an integrated hands-free sputum-processing and real-time PCR system with rapid on-demand, near-patient technology, to simultaneously detect M. tuberculosis and rifampin resistance. Analytic tests of M. tuberculosis DNA demonstrated a limit of detection (LOD) of 4.5 genomes per reaction. Studies using sputum spiked with known numbers of M. tuberculosis CFU predicted a clinical LOD of 131 CFU/ml. Killing studies showed that the assay's buffer decreased M. tuberculosis viability by at least 8 logs, substantially reducing biohazards. Tests of 23 different commonly occurring rifampin resistance mutations demonstrated that all 23 (100%) would be identified as rifampin resistant. An analysis of 20 nontuberculosis mycobacteria species confirmed high assay specificity. A small clinical validation study of 107 clinical sputum samples from suspected tuberculosis cases in Vietnam detected 29/29 (100%) smear-positive culture-positive cases and 33/39 (84.6%) or 38/53 (71.7%) smear-negative culture-positive cases, as determined by growth on solid medium or on both solid and liquid media, respectively. M. tuberculosis was not detected in 25/25 (100%) of the culture-negative samples. A study of 64 smear-positive culture-positive sputa from retreatment tuberculosis cases in Uganda detected 63/64 (98.4%) culture-positive cases and 9/9 (100%) cases of rifampin resistance. Rifampin resistance was excluded in 54/55 (98.2%) susceptible cases. Specificity rose to 100% after correcting for a conventional susceptibility test error. In conclusion, this highly sensitive and simple-to-use system can detect M. tuberculosis directly from sputum in less than 2 h.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Helb D,Jones M,Story E,Boehme C,Wallace E,Ho K,Kop J,Owens MR,Rodgers R,Banada P,Safi H,Blakemore R,Lan NT,Jones-López EC,Levi M,Burday M,Ayakaka I,Mugerwa RD,McMillan B,Winn-Deen E,Christel L,Dailey P,Perkinsdoi
10.1128/JCM.01463-09subject
Has Abstractpub_date
2010-01-01 00:00:00pages
229-37issue
1eissn
0095-1137issn
1098-660Xpii
JCM.01463-09journal_volume
48pub_type
杂志文章abstract::Thirty-three strains of Cryptococcus neoformans were isolated from clinical specimens, including specimens from AIDS patients in Brazil, and were classified into two serotypes; we detected 31 and 2 strains of serotypes A and B, respectively. Random amplified polymorphic DNA (RAPD) fingerprint pattern analyses of these...
journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.37.2.315-320.1999
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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.36.9.2439-2442.1998
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abstract::The hexagonal surface structure believed to be characteristic for influenza C virus was not initially observed for a newly isolated virus of this type. After a few additional egg passages of the virus, however, the hexagonal structure could be readily seen on many virions. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
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abstract::Several methods have been previously suggested for the presumptive identification of Gardnerella vaginalis in clinical laboratories, but none is entirely satisfactory. We previously found that sodium polyanetholesulfonate (SPS) inhibits G. vaginalis in blood culture media. We compared susceptibility to an SPS-containi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.21.2.146-149.1985
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abstract::We compared the new Abbott TestPack (TP) respiratory syncytial virus (RSV) enzyme immunoassay (EIA) with cell culture and two commercial RSV EIAs (from Abbott Diagnostics and Kallestad Laboratories) by using split samples of fresh nasal washings from children with suspected RSV disease. Two tubes of HEp-2 cells were i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.6.1395-1397.1990
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abstract::The technique of random amplified polymorphic DNA (RAPD) was adapted and optimized to study Haemophilus ducreyi isolates. A panel of 43 strains isolated from chancroid patients from different countries in Africa, Europe, North America, and Asia were characterized. The strains were also studied with respect to lipoolig...
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.37.5.1554-1560.1999
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.4.1654-1656.2001
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.6.1699-1703.1999
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.6.1355-1358.1996
更新日期:1996-06-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.4.1651-1661.2005
更新日期:2005-04-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01423-12
更新日期:2012-10-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.6.1367-1371.1989
更新日期:1989-06-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00840-12
更新日期:2012-07-01 00:00:00
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.00136-08
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.37.12.4183-4185.1999
更新日期:1999-12-01 00:00:00
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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
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doi:10.1128/JCM.35.2.477-478.1997
更新日期:1997-02-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
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abstract::New Delhi metallo-β-lactamase (NDM)-producing bacteria are considered potential global health threats. It is necessary to monitor NDM-1 and its variants in clinical isolates in order to understand the NDM-1 epidemic and the impact of its variants on β-lactam resistance. To reduce the lengthy time needed for cloning an...
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.36.9.2718-2722.1998
更新日期:1998-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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abstract::This paper describes the first isolation of a new Shigella flexneri serotype, designated 4s, in Beijing, China. Genotypic and phenotypic profiling suggests that this isolate is a clone of the S. flexneri serotype X variant reference strain. Of particular concern is the multidrug resistance exhibited by this isolate. ...
journal_title:Journal of clinical microbiology
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更新日期:2011-03-01 00:00:00
abstract::A protocol was optimized for the isolation of Mycobacterium avium subsp. paratuberculosis (MAP) from milk and colostrum, with parameters including chemical decontamination, antibiotics, and different culture media. This study demonstrates that the efficiency of MAP recovery from milk is highly dependent upon the cultu...
journal_title:Journal of clinical microbiology
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abstract::Saliva from subjects with amebic liver abscess (ALA), acute amebic colitis, asymptomatic infection with Entamoeba histolytica or Entamoeba dispar, and uninfected controls was tested by enzyme-linked immunosorbent assay (ELISA) for the presence of E. histolytica galactose-inhibitable lectin antigen and salivary immunog...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2000-06-01 00:00:00
abstract::The method of choice for the detection of Lassa virus is reverse transcription (RT)-PCR. However, the high degree of genetic variability of the virus poses a problem with the design of RT-PCR assays that will reliably detect all strains. Recently, we encountered difficulties in detecting some strains from Liberia and ...
journal_title:Journal of clinical microbiology
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abstract::A hemolysin produced by Aeromonas hydrophila CA-11, isolated from an environmental source, was purified by sulfopropyl-Sephadex C-25 chromatography at pH 5.0. This hemolysin caused fluid accumulation in infant mouse intestines and rabbit intestinal loops and killed Vero cells, as did the hemolysin produced by strain A...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.2.228-232.1986
更新日期:1986-08-01 00:00:00