Improved polyacrylamide-based artificial sputum with formalin-fixed tubercle bacilli for training of tuberculosis microscopists.

Abstract:

:Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists.

journal_name

J Clin Microbiol

authors

Yamada H,Mitarai S,Wahyunitisari MR,Mertaniasih NM,Sugamoto T,Chikamatsu K,Aono A,Matsumoto H,Fujiki A

doi

10.1128/JCM.00370-11

subject

Has Abstract

pub_date

2011-10-01 00:00:00

pages

3604-9

issue

10

eissn

0095-1137

issn

1098-660X

pii

JCM.00370-11

journal_volume

49

pub_type

杂志文章
  • Mutanolysin enhancement of serogrouping of single colonies of streptococci.

    abstract::Single colonies of beta-hemolytic streptococci could be grouped by antibody-coated latex bead agglutination or coagglutination with staphylococci coated with antibody after incubation of the colonies with mutanolysin. This simple and quick procedure provided an enzymatic means of enhancing the sensitivity of tests suc...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.15.1.184-186.1982

    authors: Calandra GB,Henson CL

    更新日期:1982-01-01 00:00:00

  • Characterization of Third-Generation-Cephalosporin-Resistant Shiga Toxin-Producing Strains of Escherichia coli O157:H7 in Japan.

    abstract::We isolated Shiga toxin-producing Escherichia coli O157:H7 strains resistant to third-generation cephalosporins. The resistant strains harbored blaCMY-2, a plasmid-mediated AmpC β-lactamase. Genotyping of isolates revealed the possible spread of this problematic bacterium. Results suggested the importance of the inves...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01263-15

    authors: Kawahara R,Seto K,Taguchi M,Nakajima C,Kumeda Y,Suzuki Y

    更新日期:2015-09-01 00:00:00

  • Culture and PCR detection of Haemophilus influenzae and Haemophilus haemolyticus in Australian Indigenous children with bronchiectasis.

    abstract::A PCR for protein D (hpd#3) was used to differentiate nontypeable Haemophilus influenzae (NTHI) from Haemophilus haemolyticus. While 90% of nasopharyngeal specimens and 100% of lower-airway specimens from 84 Indigenous Australian children with bronchiectasis had phenotypic NTHI isolates confirmed as H. influenzae, onl...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00566-12

    authors: Hare KM,Binks MJ,Grimwood K,Chang AB,Leach AJ,Smith-Vaughan H

    更新日期:2012-07-01 00:00:00

  • Diagnosis of pneumococcal pneumonia by psaA PCR analysis of lung aspirates from adult patients in Kenya.

    abstract::psaA is the gene encoding pneumococcal surface adhesin A (PsaA), a 37-kDa protein expressed on the surface of Streptococcus pneumoniae. PCR primers for psaA have been shown to amplify the target DNA sequence in all 90 serotypes of S. pneumoniae and in none of 67 heterologous pathogens and colonizing bacteria of the up...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.6.2554-2559.2003

    authors: Scott JA,Marston EL,Hall AJ,Marsh K

    更新日期:2003-06-01 00:00:00

  • Molecular characterization of pneumococcal nasopharynx isolates collected from children during their first 2 years of life.

    abstract::Pneumococcal colonization was studied in 19 children monitored from birth through the age of 2 years. For this purpose, pneumococcal isolates were characterized by capsular typing, restriction fragment end labeling (RFEL), and penicillin-binding protein (PBP) genotyping. Fifty-eight isolates were collected and were fo...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.36.8.2248-2253.1998

    authors: Sluijter M,Faden H,de Groot R,Lemmens N,Goessens WH,van Belkum A,Hermans PW

    更新日期:1998-08-01 00:00:00

  • Typing of Helicobacter pylori vacA gene and detection of cagA gene by PCR and reverse hybridization.

    abstract::The present report describes an analysis of two virulence genes of Helicobacter pylori. Parts of the cagA gene, as well as parts from the signal (s) and middle (m) regions of the mosaic vacA gene, were amplified with biotin-labelled PCR primers and the products were subsequently analyzed by a single-step reverse hybri...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.36.5.1271-1276.1998

    authors: van Doorn LJ,Figueiredo C,Rossau R,Jannes G,van Asbroek M,Sousa JC,Carneiro F,Quint WG

    更新日期:1998-05-01 00:00:00

  • Clinical evaluation of a new polymerase chain reaction assay for detection of Chlamydia trachomatis in endocervical specimens.

    abstract::A clinical evaluation of the Amplicor polymerase chain reaction (PCR) assay for the detection of Chlamydia trachomatis in endocervical swabs (Roche Molecular Systems, Branchburg, N.J.) is described. This new clinical system used one-step sample preparation, amplification with biotinylated cryptic plasmid primer pairs ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.31.10.2648-2653.1993

    authors: Bass CA,Jungkind DL,Silverman NS,Bondi JM

    更新日期:1993-10-01 00:00:00

  • Proposed changes in interpretive criteria and potency of ampicillin and ampicillin-sulbactam disks for susceptibility tests.

    abstract::The accuracy of disk susceptibility tests with ampicillin and ampicillin-sulbactam was not improved when the amount of ampicillin was increased from 10 to 20 or 30 micrograms per disk. For testing members of the family Enterobacteriaceae, ampicillin disk tests correlated better with broth microdilution tests when the ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.26.4.750-754.1988

    authors: Barry AL,Jones RN

    更新日期:1988-04-01 00:00:00

  • Predominance of the ac variant in K88-positive Escherichia coli isolates from swine.

    abstract::Monoclonal antibodies to K88ac and K88ab were used in enzyme-linked immunosorbent assays on Escherichia coli cultures known to produce K88 pili. A total of 415 K88-positive E. coli isolates from nine states were all found to be the K88ac variant. The cultures tested were isolated during the years 1976 to 1985. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.26.1.149-150.1988

    authors: Westerman RB,Mills KW,Phillips RM,Fortner GW,Greenwood JM

    更新日期:1988-01-01 00:00:00

  • Multiplex PCRs for identification of necrotoxigenic Escherichia coli.

    abstract::Two multiplex PCRs were developed for the detection of necrotoxigenic Escherichia coli virulence genes. M1 contained the primers for the toxins and the aerobactin, and M2 contained the primers for the adhesins. They were validated by single PCRs performed with reference E. coli strains and by multiplex PCRs with necro...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.9.4480-4482.2003

    authors: Van Bost S,Jacquemin E,Oswald E,Mainil J

    更新日期:2003-09-01 00:00:00

  • Immunoblot method to detect Streptococcus pneumoniae and identify multiple serotypes from nasopharyngeal secretions.

    abstract::Conventional culture techniques are limited in the ability to detect multiple Streptococcus pneumoniae serotypes in nasopharyngeal (NP) secretions. We developed an immunoblot (IB) method with monoclonal antibodies (MAbs) to detect S. pneumoniae and to identify serotypes. NP specimens stored in skim milk-tryptone-gluco...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.42.4.1596-1600.2004

    authors: Bronsdon MA,O'Brien KL,Facklam RR,Whitney CG,Schwartz B,Carlone GM

    更新日期:2004-04-01 00:00:00

  • Detection of Mycobacterium tuberculosis in BACTEC 12B broth cultures by the Roche Amplicor PCR assay.

    abstract::To evaluate the ability of the Amplicor MTB Assay to detect Mycobacterium tuberculosis complex (MTBC) organisms in BACTEC 12B broth cultures, 249 cultures with a growth index (GI) of > or = 20 from 160 patients were tested retrospectively. Specimens were processed by standard methods, and then BACTEC 12B vials and Mid...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.35.4.900-902.1997

    authors: Smith MB,Bergmann JS,Woods GL

    更新日期:1997-04-01 00:00:00

  • Synergistic lysis of erythrocytes by Propionibacterium acnes.

    abstract::Sheep and human erythrocytes, partially processed by Staphylococcus aureus or Clostridium perfringens, were susceptible to lysis in the presence of Propionibacterium acnes. P. acnes liberated a lipase that was detected on Tween 80 agar and also on phospholipase C-precipitated egg yolk agar. Such a lipase might have co...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Choudhury TK

    更新日期:1978-08-01 00:00:00

  • Assessment of Etest as an alternative to agar dilution for antimicrobial susceptibility testing of Neisseria gonorrhoeae.

    abstract::We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory I...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02131-13

    authors: Liu H,Taylor TH Jr,Pettus K,Trees D

    更新日期:2014-05-01 00:00:00

  • Identification of epidemic strains of Acinetobacter baumannii by integrase gene PCR.

    abstract::Forty-eight clinical Acinetobacter isolates with different epidemic behavior were investigated for the presence of integrons and plasmids and for antibiotic susceptibility. Integrons were demonstrated in 50% of the strains by an integrase gene PCR. Epidemic strains of Acinetobacter baumannii were found to contain sign...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.39.1.8-13.2001

    authors: Koeleman JG,Stoof J,Van Der Bijl MW,Vandenbroucke-Grauls CM,Savelkoul PH

    更新日期:2001-01-01 00:00:00

  • High rate of tetracycline resistance in Streptococcus pyogenes in Iran: an epidemiological study.

    abstract::Streptococcus pyogenes, a major human pathogen, is still considered susceptible to beta-lactams, but for other relevant antibiotics, highly variable resistance rates have been reported. Since no data were available from Iran, we tested 1,335 throat isolates from two different regions of the country for their antibioti...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Jasir A,Tanna A,Noorani A,Mirsalehian A,Efstratiou A,Schalen C

    更新日期:2000-06-01 00:00:00

  • Rapid flow cytometric susceptibility testing of Candida albicans.

    abstract::A rapid flow cytometric assay for in vitro antifungal drug susceptibility testing was developed by adapting the proposed reference method for broth macrodilution testing of yeasts. Membrane permeability changes caused by the antifungal agent were measured by flow cytometry using propidium iodide, a nucleic acid-bindin...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.35.9.2320-2324.1997

    authors: Ramani R,Ramani A,Wong SJ

    更新日期:1997-09-01 00:00:00

  • Compliance of clinical microbiology laboratories in the United States with current recommendations for processing respiratory tract specimens from patients with cystic fibrosis.

    abstract::Respiratory tract specimens from patients with cystic fibrosis (CF) require unique processing by clinical microbiology laboratories to ensure detection of all potential pathogens. The present study sought to determine the compliance of microbiology laboratories in the United States with recently published recommendati...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.44.4.1547-1549.2006

    authors: Zhou J,Garber E,Desai M,Saiman L

    更新日期:2006-04-01 00:00:00

  • Visuwell Reagin, a non-treponemal enzyme-linked immunosorbent assay for the serodiagnosis of syphilis.

    abstract::A urease-based enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of reagin antibodies in serum. Visuwell Reagin (ADI Diagnostics Inc., Rexdale, Ontario, Canada) is a non-treponemal screening test for the serodiagnosis of syphilis which has the benefits of large batch testing, automatabilit...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.27.10.2300-2304.1989

    authors: White TJ,Fuller SA

    更新日期:1989-10-01 00:00:00

  • Detection of duck hepatitis B virus DNA on filter paper by PCR and SYBR green dye-based quantitative PCR.

    abstract::Duck hepatitis B virus (DHBV) belongs to the Hepadnaviridae family, which includes human Hepatitis B virus (HBV) and Woodchuck hepatitis virus. It is widely distributed in wild and domestic ducks due to congenital transmission. HBV is a worldwide health problem, with carriers at risk of developing cirrhosis and liver ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.40.7.2584-2590.2002

    authors: Wang CY,Giambrone JJ,Smith BF

    更新日期:2002-07-01 00:00:00

  • Dichotomy of glycoprotein g gene in herpes simplex virus type 1 isolates.

    abstract::Herpes simplex virus type 1 (HSV-1) encodes 11 envelope glycoproteins, of which glycoprotein G-1 (gG-1) induces a type-specific antibody response. Variability of the gG-1 gene among wild-type strains may be a factor of importance for a reliable serodiagnosis and typing of HSV-1 isolates. Here, we used a gG-1 type-spec...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.40.9.3245-3251.2002

    authors: Rekabdar E,Tunbäck P,Liljeqvist JA,Lindh M,Bergström T

    更新日期:2002-09-01 00:00:00

  • Characterization of rough and smooth morphotypes of Mycobacterium abscessus isolates from clinical specimens.

    abstract::Mycobacterium abscessus, which consists of the two subspecies M. abscessus subspecies abscessus and M. abscessus subspecies bolletii, can produce rough or smooth colony morphologies. Here we analyzed 50 M. abscessus isolates cultured from the respiratory specimens of 34 patients, 28 (82%) of whom had cystic fibrosis (...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01249-13

    authors: Rüger K,Hampel A,Billig S,Rücker N,Suerbaum S,Bange FC

    更新日期:2014-01-01 00:00:00

  • Evaluation of an enzyme-linked immunosorbent assay for the detection of ectromelia (mousepox) antibody.

    abstract::Ectromelia virus, an orthopoxvirus that can cause extensive morbidity and mortality (mousepox) in colonized mice, has been epizootically responsible for serious disruption of biomedical research since 1930. The lack of a sensitive and specific serological assay for infection with this virus became apparent during outb...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.18.5.1220-1225.1983

    authors: Buller RM,Bhatt PN,Wallace GD

    更新日期:1983-11-01 00:00:00

  • Worldwide evaluation of DNA sequencing approaches for identification of drug resistance mutations in the human immunodeficiency virus type 1 reverse transcriptase.

    abstract::A panel (ENVA-1) of well-defined blinded samples containing wild-type and mutant human immunodeficiency virus type 1 (HIV-1) reverse transcriptase was analyzed by automated DNA sequencing in 23 laboratories worldwide. Drug resistance mutations at codons 41, 215, and 184 were present in the panel samples at different r...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章,多中心研究

    doi:10.1128/JCM.37.7.2291-2296.1999

    authors: Schuurman R,Demeter L,Reichelderfer P,Tijnagel J,de Groot T,Boucher C

    更新日期:1999-07-01 00:00:00

  • Rapid identification of bacterial pathogens in positive blood culture bottles by use of a broad-based PCR assay coupled with high-resolution melt analysis.

    abstract::We evaluated a broad-based PCR assay coupled with high-resolution melt analysis for rapid bacterial identification in patients with bacterial sepsis. With a reference library of 60 clinically relevant bacterial species, 52 positive blood culture samples were tested. Our assay identified 46/52 samples at the species le...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00718-10

    authors: Won H,Rothman R,Ramachandran P,Hsieh YH,Kecojevic A,Carroll KC,Aird D,Gaydos C,Yang S

    更新日期:2010-09-01 00:00:00

  • Assessment on experimental bacterial biofilms and in clinical practice of the efficacy of sampling solutions for microbiological testing of endoscopes.

    abstract::Opinions differ on the value of microbiological testing of endoscopes, which varies according to the technique used. We compared the efficacy on bacterial biofilms of sampling solutions used for the surveillance of the contamination of endoscope channels. To compare efficacy, we used an experimental model of a 48-h Ps...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.06221-11

    authors: Aumeran C,Thibert E,Chapelle FA,Hennequin C,Lesens O,Traoré O

    更新日期:2012-03-01 00:00:00

  • PCR strategy for identification and differentiation of small pox and other orthopoxviruses.

    abstract::Rapid identification and differentiation of orthopoxviruses by PCR were achieved with primers based on genome sequences encoding the hemagglutinin (HA) protein, an infected-cell membrane antigen that distinguishes orthopoxviruses from other poxvirus genera. The initial identification step used a primer pair of consens...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.8.2069-2076.1995

    authors: Ropp SL,Jin Q,Knight JC,Massung RF,Esposito JJ

    更新日期:1995-08-01 00:00:00

  • Comparison of plasmid profiles, phage types, and antimicrobial resistance patterns of Salmonella enteritidis isolates in the United States.

    abstract::To evaluate the laboratory techniques for subtyping isolates of Salmonella enteritidis, we compared the plasmid profiles (PP), phage types (PT), and antimicrobial susceptibility patterns (AS) of two nationally representative samples of sporadic human S. enteritidis isolates from 1979 (n = 28) and 1984 (n = 37), 43 iso...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.30.4.854-857.1992

    authors: Rodrigue DC,Cameron DN,Puhr ND,Brenner FW,St Louis ME,Wachsmuth IK,Tauxe RV

    更新日期:1992-04-01 00:00:00

  • Combination assay detecting both human immunodeficiency virus (HIV) p24 antigen and anti-HIV antibodies opens a second diagnostic window.

    abstract::Fourth-generation human immunodeficiency virus (HIV) screening immunoassays reduce the diagnostic window between infection and diagnosis by the inclusion of HIV p24 antigen detection together with HIV antibody detection in the same test. We compared third- and fourth-generation HIV immunoassays and a dedicated HIV p24...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.43.10.5397-5399.2005

    authors: Speers D,Phillips P,Dyer J

    更新日期:2005-10-01 00:00:00

  • Electrophoretic karyotyping of typical and atypical Candida albicans.

    abstract::Electrophoretic karyotypes of atypical isolates of Candida albicans, e.g., strains that were germ tube negative, failed to express proteinase activity, demonstrated low virulence for mice, formed hyperchlamydoconidia, produced hyperhyphae, or were sucrose negative (including the type strain of Candida stellatoidea), w...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.5.876-881.1990

    authors: Mahrous M,Lott TJ,Meyer SA,Sawant AD,Ahearn DG

    更新日期:1990-05-01 00:00:00