Abstract:
:Recent outbreaks of severe acute respiratory syndrome (SARS) have spurred intense research efforts around the world to deal with the serious threat to health posed by this novel coronavirus. A rapid, reliable diagnostic assay is needed for monitoring the spread of the disease. Here we report a method for eliminating false-negative results and increasing test sensitivity, based on the hypothesis that the message encoded by the nucleocapsid (N) gene is the most abundant during viral infection. Nasopharyngeal aspirates and stool samples were obtained from suspected SARS patients with major clinical symptoms and a significant history of close contact with infected patients. Total RNAs were extracted in a 96-well format, together with pig kidney epithelial (PK-15) cells as an internal control for extraction efficiency. PCR inhibitors were removed by ethanol precipitation, and a PCR for the pig beta-actin gene was used as a positive control for all clinical samples. Samples were analyzed by a reverse transcriptase PCR assay. Northern blot analysis was performed to demonstrate differences in subgenomic transcripts of the virus, and a real-time quantitative PCR was employed to compare the sensitivities of two loci (1b and N). The detection rate of the assay reached 44.4% on day 9 after the onset of the disease. The diagnostic PCR amplifying the N gene gave an average of a 26.0% (6.3 to 60.0%) stronger intensity signal than that for the 1b gene. In conclusion, the nucleocapsid gene represents an additional sensitive molecular marker for the diagnosis of the SARS coronavirus and can be further adapted for use in a high-throughput platform assay.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Hui RK,Zeng F,Chan CM,Yuen KY,Peiris JS,Leung FCdoi
10.1128/jcm.42.5.1994-1999.2004subject
Has Abstractpub_date
2004-05-01 00:00:00pages
1994-9issue
5eissn
0095-1137issn
1098-660Xjournal_volume
42pub_type
杂志文章abstract::Bordetella pertussis was transmitted from an immunized boy to his father and possibly to other family members. This case report demonstrates that although unusual, acute adult B. pertussis infection can occur. B. pertussis immunization may not prevent infection but can reduce the severity. B. pertussis was detected in...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.2.429-430.1996
更新日期:1996-02-01 00:00:00
abstract::A total of 215 typical strains of Candida albicans were studied for their ability to assimilate 11 carbon compounds. All isolates assimilated lactic acid, ribitol, succinic acid, methyl alpha-D-glucopyranoside. None of the isolates assimilated cellobiose and salicin; 1.9% of the isolates assimilated L-arabinose. Citri...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.13.1.163-166.1981
更新日期:1981-01-01 00:00:00
abstract::The duration of mumps virus RNA detection was studied during a mumps outbreak in a highly vaccinated university population. Seven of the eight reverse transcription-PCR-positive specimens were collected during the first 3 days of parotitis, suggesting that viral shedding is minimal after the first 3 days of symptoms. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01803-07
更新日期:2008-03-01 00:00:00
abstract::In anticipation of rotavirus vaccine introduction in Saudi Arabia, this study was undertaken to determine the distribution of the G and P genotypes of rotaviruses in order to examine whether there was any emerging serotype or unusual strain circulating in children in Saudi Arabia. Of 984 stool specimens collected betw...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02244-07
更新日期:2008-04-01 00:00:00
abstract::An association between (unculturable) gastrospirillum-like organisms (GLO) and ulcerative lesions in the pars oesophagea in stomachs of swine has been claimed. In dogs GLO detected by microscopy may represent several Helicobacter species or subspecies. Therefore we investigated which Helicobacter spp. are present in s...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.7.2661-2664.2000
更新日期:2000-07-01 00:00:00
abstract::The new anaerobe and Corynebacterium (ANC) identification card for Vitek 2 was compared with a 16S rRNA gene sequencing (16S) reference method for accuracy in the identification of corynebacteria and anaerobic species. Testing was performed on a Vitek 2 XL system with modified software at three clinical trial laborato...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.00450-08
更新日期:2008-08-01 00:00:00
abstract::Using the Staphylococcus aureus absorption method, we have modified a commercial radioimmunoassay kit for the determination of total antibody to hepatitis B core antigen so that we can now determine the predominant globulin species of that antibody. One-hundred percent of acute hepatitis B sera had a predominance of i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.16.5.973-975.1982
更新日期:1982-11-01 00:00:00
abstract::During pregnancy, Plasmodium falciparum infection of the placenta frequently occurs in the absence of parasites in peripheral blood. We investigated the abilities of the OptiMAL rapid immunochromatographic strip test for P. falciparum lactate dehydrogenase and species-specific PCR performed on peripheral blood to dete...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.1.155-158.2002
更新日期:2002-01-01 00:00:00
abstract::Transfusion-acquired cytomegalovirus (CMV) infections should be prevented in seronegative immunocompromised patients by providing blood products from donors who are also seronegative. Latex agglutination was investigated as a simple and rapid method for detecting antibody against CMV. Latex beads were coated with CMV ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.22.1.68-70.1985
更新日期:1985-07-01 00:00:00
abstract::The mumps virus is a negative-strand RNA virus in the family Paramyxoviridae. Mumps infection results in an acute illness with symptoms including fever, headache, and myalgia, followed by swelling of the salivary glands. Complications of mumps can include meningitis, deafness, pancreatitis, orchitis, and first-trimest...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00614-07
更新日期:2007-09-01 00:00:00
abstract::Direct tube coagulase testing for identification of Staphylococcus aureus from BACTEC culture broth showed a sensitivity, a specificity, and positive and negative predicative values of 34%, 100%, 100%, and 80.2% with 2 h of incubation and 65%, 98.7, 99.7%, and 88.6% with 4 h of incubation. Anaerobic blood culture cont...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00369-07
更新日期:2007-07-01 00:00:00
abstract::We reviewed laboratory-acquired infections occurring in Utah from 1978 through 1982. Written and telephone interviews of supervisors of 1,191 laboratorians revealed an estimated annual incidence of 3 laboratory-acquired infections per 1,000 employees. Infections, in order of frequency, included hepatitis B (clinical c...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.21.4.486-489.1985
更新日期:1985-04-01 00:00:00
abstract::Cowan I staphylococci coated with antisera to streptococcal groups A, B, C, D, F, and G were used as coagglutination reagents in a modified coagglutination procedure (MCAP). Streptococcal group antigens were extracted with a Streptomyces albus-lysozyme enzyme mixture for 30 min at 55 degrees C and centrifuged, and the...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1979-03-01 00:00:00
abstract::Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular epidemiological study. Repetitive extragenic palindromic PCR (rep-PCR) is less time-consuming and more suitable for analyzing large numbers of bacterial strains in human populations. PFGE and rep-PCR provide comparable genotyping r...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01828-09
更新日期:2010-02-01 00:00:00
abstract::Widespread use of conjugate pneumococcal polysaccharide-protein vaccines may alter the spectrum of pneumococci producing invasive disease. Novel sensitive diagnostic methods would be valuable for monitoring the epidemiology of pneumococcal disease within populations and vaccine recipients. Ideally, these methods shoul...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.10.4972-4976.2005
更新日期:2005-10-01 00:00:00
abstract::Seventy-eight blood cultures with a Gram stain result of Gram-positive cocci in pairs and/or chains were evaluated with the Nanosphere Verigene Gram-positive blood culture (BC-GP) assay. The overall concordance of the assay with culture was 89.7% (70/78 cultures), allowing for the development of a targeted treatment a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01587-13
更新日期:2013-11-01 00:00:00
abstract::Parasite-specific humoral and cellular immune responses were evaluated in in nine immunosuppressed patients with Strongyloides stercoralis infection. Four patients had the disseminated form of the disease, and five had serious manifestations without evidence of hyperinfection. All patients had high levels of immunoglo...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.6.1099-1103.1986
更新日期:1986-06-01 00:00:00
abstract::The past 2 decades have brought worrying increases in severe Streptococcus pyogenes diseases globally. To investigate and compare the epidemiological patterns of these diseases within Europe, data were collected through a European Union FP-5-funded program (Strep-EURO). Prospective population-based surveillance of sev...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00422-08
更新日期:2008-07-01 00:00:00
abstract::In Asia, bla(KPC) detection has been limited to East Asia and not yet seen in Southeast Asia. We report four bla(KPC-2)-containing Klebsiella pneumoniae isolates from two different hospitals in Singapore. All isolates belonged to strain type 11 (ST11) and were indistinguishable by pulsed-field gel electrophoresis (PFG...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.05914-11
更新日期:2012-02-01 00:00:00
abstract::To differentiate the Borrelia burgdorferi sensu lato genospecies, LightCycler real-time PCR was used for the fluorescence (SYBR Green I) melting curve analysis of borrelial recA gene PCR products. The specific melting temperature analyzed is a function of the GC/AT ratio, length, and nucleotide sequence of the amplifi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.7.2756-2759.2000
更新日期:2000-07-01 00:00:00
abstract::Tubercle bacilli may survive in unstained heat-fixed sputum smears and may be an infection risk to laboratory staff. We compared the effectiveness of 1% and 5% sodium hypochlorite, 5% phenol, 2% glutaraldehyde, and 3.7% formalin in killing Mycobacterium tuberculosis present in smears prepared from 51 sputum samples. T...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.8.4245-4248.2005
更新日期:2005-08-01 00:00:00
abstract::A comparative study of herpes simplex virus diagnosis by standard cell culture and a new hybrid test (enzyme-linked immunosorbent assay spin amplification technique) was done on 300 specimens. The new test was found to be equally sensitive and specific, much less expensive to perform, and to report all results in 48 h...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.2.310-311.1986
更新日期:1986-08-01 00:00:00
abstract::In Brazil, the most common etiological agent of American tegumentary leishmaniasis is Leishmania (Viannia) braziliensis. In general, diagnostic techniques envisage the visualization of the parasite, but that technique has a low sensitivity. The main purpose of the present work was to evaluate the PCR as a routine tool...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.6.1819-1823.1999
更新日期:1999-06-01 00:00:00
abstract::A PCR test based on insertion sequence IS1081 was developed to detect Mycobacterium tuberculosis complex organisms in the peripheral blood. The method was applied to blood samples from immunocompetent individuals with localized pulmonary tuberculosis. Seven of 16 (43.75%) blood samples were found to be positive for th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.10.3094-3095.1998
更新日期:1998-10-01 00:00:00
abstract::A member of the Enterobacteriaceae initially identified as Kluyvera cryocrescens by the MicroScan Gram-Negative Combo 13 panel caused an outbreak of nosocomial infections in four patients (pneumonia, n = 2; urinary tract infection, n = 1; wound infection, n = 1) and urinary tract colonization in one patient. When the ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.11.3946-3952.2000
更新日期:2000-11-01 00:00:00
abstract::Antigens of Rhodococcus equi were analyzed by immunoblotting with naturally infected foal sera. Immunoblots of whole-cell antigen preparations of clinical isolates of R. equi revealed that major protein bands with molecular masses of 15 to 17 kDa were present in all clinical isolates tested and all isolates virulent f...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.3.439-443.1991
更新日期:1991-03-01 00:00:00
abstract::Over a 6-year period, 24 extended-spectrum beta-lactamase (ESBL)-producing isolates of Pseudomonas aeruginosa were collected from 18 patients living in a nursing home. These isolates had a delayed development of a red pigment and exhibited a similar antibiotype (resistance to all beta-lactams except for imipenem and t...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.8.4129-4138.2005
更新日期:2005-08-01 00:00:00
abstract::An indirect fluorescent antibody technique has been developed for the serological diagnosis of gonorrhea. The selected strain(s) of Neisseria gonorrhoeae possesses a heat-labile surface antigen (L-antigen). Sera are diluted 1:10, and an aliquot is hear inactivated at 59 C for 30 min. The treated and untreated aliquots...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1976-04-01 00:00:00
abstract::Six isolates of Mycobacterium avium of genotype dnaJ(+) IS901(-) IS1311(+) IS1245(+) and serotypes 6 (n = 1), 6/9, (n = 2), and 9 (n = 3) were obtained within a 5-month period from a human immunodeficiency virus-negative patient treated for tuberculosis. The isolates were identified with PvuII restriction fragment len...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.10.3712-3719.2002
更新日期:2002-10-01 00:00:00
abstract::A combination of immunomagnetic separation (IMS) and a polymerase chain reaction (PCR) procedure was used for direct isolation and identification of Shigella dysenteriae type 1 and Shigella flexneri from feces. Immunomagnetic particles were coated with monoclonal antibody MASFB, which is specific for a common epitope ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.11.2801-2806.1992
更新日期:1992-11-01 00:00:00