Single-step multiplex PCR assay for characterization of New World Leishmania complexes.

Abstract:

:We have developed a PCR assay for one-step differentiation of the three complexes of New World Leishmania (Leishmania braziliensis, Leishmania mexicana, and Leishmania donovani). This multiplex assay is targeted to the spliced leader RNA (mini-exon) gene repeats of these organisms and can detect all three complexes simultaneously, generating differently sized products for each complex. The assay is specific to the Leishmania genus and does not recognize related kinetoplastid protozoa, such as Trypanosoma cruzi, Trypanosoma brucei, and Crithidia fasciculata. It correctly identified Leishmania species with a broad geographic distribution in Central and South America. The sensitivity of the PCR amplification ranged from 1 fg to 10 pg of DNA (0.01 to 100 parasites), depending on the complex detected. Crude extracts of cultured parasites, prepared simply by boiling diluted cultures, served as excellent templates for amplification. Crude preparations of clinical material were also tested. The assay detected L. braziliensis in dermal scrapings from cutaneous leishmanial lesions, Leishmania chagasi in dermal scrapings of atypical cutaneous leishmaniasis, and L. mexicana from lesion aspirates from infected hamsters. We have minimized the material requirements and maximized the simplicity, rapidity, and informative content of this assay to render it suitable for use in laboratories in countries where leishmaniasis is endemic. This assay should be useful for rapid in-country identification of Leishmania parasites, particularly where different Leishmania complexes are found in the same geographical area.

journal_name

J Clin Microbiol

authors

Harris E,Kropp G,Belli A,Rodriguez B,Agabian N

doi

10.1128/JCM.36.7.1989-1995.1998

subject

Has Abstract

pub_date

1998-07-01 00:00:00

pages

1989-95

issue

7

eissn

0095-1137

issn

1098-660X

journal_volume

36

pub_type

杂志文章
  • Molecular quantification of respiratory syncytial virus in respiratory samples: reliable detection during the initial phase of infection.

    abstract::Quantitative real-time PCR for the detection of respiratory syncytial virus (RSV) RNA is increasingly used to study the causal role of RSV in lower airway disease. The objective of our study was to evaluate variations in RSV RNA loads at different steps in the RNA quantification process: (i) variation in RSV RNA load ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00097-10

    authors: van de Pol AC,Wolfs TF,van Loon AM,Tacke CE,Viveen MC,Jansen NJ,Kimpen JL,Rossen JW,Coenjaerts FE

    更新日期:2010-10-01 00:00:00

  • Slaughter pigs are commonly infected by closely related but distinct gastric ulcerative lesion-inducing gastrospirilla.

    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

    authors: Roosendaal R,Vos JH,Roumen T,van Vugt R,Cattoli G,Bart A,Klaasen HL,Kuipers EJ,Vandenbroucke-Grauls CM,Kusters JG

    更新日期:2000-07-01 00:00:00

  • Taxonomic subgroups of Pasteurella multocida correlate with clinical presentation.

    abstract::Pasteurella multocida is a small nonmotile gram-negative coccobacillus that is found in the nasopharynx and gastrointestinal tract of many wild and domesticated animals. In humans it most commonly causes cellulitis and localized superficial skin abscesses following an animal bite or scratch. The respiratory tract is t...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.40.9.3438-3441.2002

    authors: Chen HI,Hulten K,Clarridge JE 3rd

    更新日期:2002-09-01 00:00:00

  • Rapid detection of Mycobacterium tuberculosis in contaminated BACTEC 12B broth cultures by testing with Amplified Mycobacterium Tuberculosis Direct Test.

    abstract::Contamination of broth cultures of acid-fast bacilli (AFB) by bacterial species other than Mycobacterium species frequently occurs. Many of these contaminated cultures require redecontamination and reincubation before the appropriate tests can be performed for identification, significantly affecting the turnaround tim...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.39.10.3718-3720.2001

    authors: Zheng X,Pang M,Engler HD,Tanaka S,Reppun T

    更新日期:2001-10-01 00:00:00

  • Bacteremia associated with Haemophilus influenzae type e biotype 4.

    abstract::Haemophilus influenzae type e biotype 4 was isolated from a single antemortem blood culture obtained from a 60-year-old, white male with abdominal carcinomatosis. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Cawthern TH,Marraro RV,McCleskey FK

    更新日期:1978-01-01 00:00:00

  • Simple, sensitive, and specific detection of human immunodeficiency virus type 1 in clinical specimens by polymerase chain reaction with nested primers.

    abstract::A simple, sensitive, and specific polymerase chain reaction (PCR) protocol for the detection of human immunodeficiency virus type 1 (HIV-1) is described. We have improved all three PCR steps: sample preparation, DNA amplification, and detection of the amplified product. Some of the improvements have been described pre...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.28.7.1560-1564.1990

    authors: Albert J,Fenyö EM

    更新日期:1990-07-01 00:00:00

  • Detection and identification of Mycobacterium spp. in clinical specimens by combining the Roche Cobas Amplicor Mycobacterium tuberculosis assay with Mycobacterium genus detection and nucleic acid sequencing.

    abstract::We have recently developed a PCR assay for detection of Mycobacterium spp. at the genus level based on the Cobas Amplicor platform. The sensitivities for smear-positive and smear-negative specimens were found to be 100% and 47.9%, respectively. The specificity was 97.7%, the positive predictive value 84.6%, and the ne...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00851-10

    authors: Peter-Getzlaff S,Lüthy J,Voit A,Bloemberg GV,Böttger EC

    更新日期:2010-11-01 00:00:00

  • Three variants of Corynebacterium diphtheriae subsp. mitis (Belfanti) isolated from a throat specimen.

    abstract::Three variants of a Corynebacterium diphtheriae subsp. mitis strain belfanti, two toxigenic (phage types 7 and 11) and one nontoxigenic (phage type 7), were simultaneously isolated from the throat of an adult. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Chang DN,Laughren GS,Chalvardjian NE

    更新日期:1978-12-01 00:00:00

  • Epidemiology and clinical presentations of the four human coronaviruses 229E, HKU1, NL63, and OC43 detected over 3 years using a novel multiplex real-time PCR method.

    abstract::Four human coronaviruses (HCoV-229E, HCoV-HKU1, HCoV-NL63, and HCoV-OC43) are associated with a range of respiratory outcomes, including bronchiolitis and pneumonia. Their epidemiologies and clinical characteristics are poorly described and are often reliant on case reports. To address these problems, we conducted a l...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00636-10

    authors: Gaunt ER,Hardie A,Claas EC,Simmonds P,Templeton KE

    更新日期:2010-08-01 00:00:00

  • Selenomonas sputigena septicemia.

    abstract::Selenomonas sputigena is a curved motile anaerobic gram-negative rod that is a part of the normal upper respiratory tract of man. We present a case of speticemia associated with this organism and believe it to be the first such report describing systemic disease. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.14.6.684-685.1981

    authors: McCarthy LR,Carlson JR

    更新日期:1981-12-01 00:00:00

  • Unique p24 epitope marker to identify multiple human immunodeficiency virus variants in blood from the same individuals.

    abstract::The human immunodeficiency virus (HIV) was isolated from the blood of 192 of 410 seropositive individuals. Original isolations were made in peripheral blood mononuclear cell (PBMC) cultures, and only one-fifth of the HIV isolates could be adapted to replicate in continuous T-cell lines. Of the 192 HIV isolates, 42 had...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.25.8.1411-1415.1987

    authors: Bolton V,Pedersen NC,Higgins J,Jennings M,Carlson J

    更新日期:1987-08-01 00:00:00

  • Recovery of Histoplasma capsulatum from blood in a commercial radiometric Mycobacterium medium.

    abstract::We report the recovery of Histoplasma capsulatum from blood specimens cultured for Mycobacterium sp. in BACTEC 13A radiometric medium. H. capsulatum was recovered from six of eight blood specimens submitted for mycobacterial cultures from five human immunodeficiency virus-positive individuals. Initial positive metabol...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.30.1.237-239.1992

    authors: Merz WG,Kodsy S,Merz CS

    更新日期:1992-01-01 00:00:00

  • Diagnosis of influenza A virus infections by detection of specific immunoglobulins M, A, and G in serum.

    abstract::The value of immunoglobulin M (IgM) detection in the early diagnosis of influenza A was examined in a prospective study during an outbreak of influenza A/Philippines/2/82 (H3N2) virus infection in February and March 1986. The IgM response was investigated in sera from 64 adults with influenzalike symptoms; we found a ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.27.3.453-458.1989

    authors: Vikerfors T,Lindegren G,Grandien M,van der Logt J

    更新日期:1989-03-01 00:00:00

  • Prediction of decreased susceptibility to penicillin of Neisseria meningitidis strains by real-time PCR.

    abstract::Sequence analysis of the penA gene, encoding penicillin-binding protein 2 (PBP2), in 30 penicillin-intermediate (PenI) Neisseria meningitidis strains showed altered gene sequences due to the translocation of exogenous DNA blocks derived from commensal neisseriae, which are known to have PBP2 proteins with decreased af...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.10.4666-4670.2003

    authors: Stefanelli P,Carattoli A,Neri A,Fazio C,Mastrantonio P

    更新日期:2003-10-01 00:00:00

  • Role of a cefoxitin-inducible beta-lactamase in a case of breakthrough bacteremia.

    abstract::Development of resistance during therapy with cefamandole contributes to treatment failure. A simple cefoxitin disk test was recently described which detects a cefamandole-active inducible beta-lactamase not otherwise detectable with cefamandole as the inducer. A case of breakthrough Enterobacter bacteremia due to se...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.12.4.517-520.1980

    authors: Beckwith DG,Jahre JA

    更新日期:1980-10-01 00:00:00

  • Association of putative enteroaggregative Escherichia coli virulence genes and biofilm production in isolates from travelers to developing countries.

    abstract::Enteroaggregative Escherichia coli (EAEC) is an emerging enteric pathogen that causes acute and chronic diarrhea among children, human immunodeficiency virus-infected patients, and travelers to developing regions of the world. The pathogenesis of EAEC strains involves the production of biofilm. In this study, we deter...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01128-06

    authors: Mohamed JA,Huang DB,Jiang ZD,DuPont HL,Nataro JP,Belkind-Gerson J,Okhuysen PC

    更新日期:2007-01-01 00:00:00

  • Comparison of Counterimmunoelectrophoresis and electron microscopy for laboratory diagnosis of human reovirus-like agent-associated infantile gastroenteritis.

    abstract::Counterimmunoelectrophoresis was compared with electron microscopy for detection of human reovirus-like agent in fecal specimens. Both tests gave very similar results. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Spence L,Fauvel M,Petro R,Bloch S

    更新日期:1977-02-01 00:00:00

  • Studies on neutralization of human serum bactericidal activity by sodium amylosulfate.

    abstract::The synthetic anticoagulant sodium amylosulfate (SAS) at concentrations of 125 to 2,000 microgram/ml failed to completely neutralize the bactericidal activity of 80 and 50% (by volume) fresh human serum. Furthermore, SAS failed to inhibit the alternative pathway of complement activation in 80% (by volume) fresh human ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Traub WH

    更新日期:1977-08-01 00:00:00

  • Frequency of Positive Aspergillus Tests in COVID-19 Patients in Comparison to Other Patients with Pulmonary Infections Admitted to the ICU.

    abstract::The aim of this study was to describe the frequency of positive Aspergillus tests in COVID-19 patients and investigate the association between COVID-19 and a positive Aspergillus test result. We compared the proportion of positive Aspergillus tests in COVID-19 patients admitted to the ICU for > 24 hours with two contr...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02278-20

    authors: Yusuf E,Vonk A,van den Akker JPC,Bode L,Sips GJ,Rijnders BJA,de Steenwinkel J,Verkaik NJ,Vogel M,van der Eerden M,van Westreenen M

    更新日期:2020-12-04 00:00:00

  • Reduced virulence of Pseudomonas aeruginosa grown in the presence of benzalkonium chloride.

    abstract::Resistant cells of Pseudomonas aeruginosa ATCC 9027 which were grown in the presence of 1 mg of benzalkonium chloride (BC) per ml caused only a mild conjunctivitis when they were dropped onto the scratched corneas of rabbits. In contrast, cells of the BC-sensitive parent strain induced a severe keratoconjunctivitis. I...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.1.2.175-179.1975

    authors: Adair FW,Liauw HL,Geftic SG,Gelzer J

    更新日期:1975-02-01 00:00:00

  • Standardization of Operator-Dependent Variables Affecting Precision and Accuracy of the Disk Diffusion Method for Antibiotic Susceptibility Testing.

    abstract::Parameters like zone reading, inoculum density, and plate streaking influence the precision and accuracy of disk diffusion antibiotic susceptibility testing (AST). While improved reading precision has been demonstrated using automated imaging systems, standardization of the inoculum and of plate streaking have not bee...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02351-15

    authors: Hombach M,Maurer FP,Pfiffner T,Böttger EC,Furrer R

    更新日期:2015-12-01 00:00:00

  • Rapid indirect hemagglutination test for serotyping Pasteurella haemolytica.

    abstract::A modified indirect hemagglutination test which enabled Pasteurella haemolytica to be serotyped in 3 h is described. There was 100% correlation with the conventional test for both the 15 prototype strains and the 50 field isolates. It was found that nonviable cultures could also be serotyped. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.18.1.206-207.1983

    authors: Fraser J,Donachie W,Quirie M,Gilmour NJ

    更新日期:1983-07-01 00:00:00

  • Typing of human enterovirus by partial sequencing of VP2.

    abstract::The sequencing of the VP1 hypervariable region of the human enterovirus (HEV) genome has become the reference test for typing field isolates. This study describes a new strategy for typing HEV at the serotype level that uses a reverse transcription-PCR assay targeting the central part of the VP2 capsid protein. Two pa...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00093-07

    authors: Nasri D,Bouslama L,Omar S,Saoudin H,Bourlet T,Aouni M,Pozzetto B,Pillet S

    更新日期:2007-08-01 00:00:00

  • Characterization of toxigenic Corynebacterium ulcerans strains isolated from humans and domestic cats in the United Kingdom.

    abstract::In the United Kingdom there has been a marked increase in the number of human infections caused by toxigenic Corynebacterium ulcerans. During 2002 and 2003 the organism was also isolated from several domestic cats with bilateral nasal discharge. As C. ulcerans has never previously been isolated from cats, the 16S rRNA...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.43.9.4377-4381.2005

    authors: De Zoysa A,Hawkey PM,Engler K,George R,Mann G,Reilly W,Taylor D,Efstratiou A

    更新日期:2005-09-01 00:00:00

  • Web-accessible database of hsp65 sequences from Mycobacterium reference strains.

    abstract::Mycobacteria include a large number of pathogens. Identification to species level is important for diagnoses and treatments. Here, we report the development of a Web-accessible database of the hsp65 locus sequences (http://msis.mycobacteria.info) from 149 out of 150 Mycobacterium species/subspecies. This database can ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02602-10

    authors: Dai J,Chen Y,Lauzardo M

    更新日期:2011-06-01 00:00:00

  • Differentiation of Bacteroides fragilis species by gas chromatographic detection of phenylacetic acid.

    abstract::Of 382 strains tested, 261 strains of the species Bacteroides fragilis, B. thetaiotaomicron, B. ovatus, and B. distasonis produced phenylacetic acid; the remaining strains, belonging exclusively to the species B. vulgatus, failed to do so. This differentiation characteristic may be useful in routine clinical bacteriol...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: van Assche PF

    更新日期:1978-11-01 00:00:00

  • Methicillin-Resistant Staphylococcus aureus spa Type t002 Outbreak in Horses and Staff at a Veterinary Teaching Hospital after Its Presumed Introduction by a Veterinarian.

    abstract::Methicillin-resistant Staphylococcus aureus (MRSA) infection and colonization, involving MRSA strains which differ from common human health care-associated clones, have become serious emerging conditions in equine veterinary hospitals. In 2010, MRSA spa type t535 caused an outbreak involving both horses and personnel ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00090-15

    authors: Steinman A,Masarwa S,Tirosh-Levy S,Gleser D,Kelmer G,Adler A,Carmeli Y,Schwaber MJ

    更新日期:2015-09-01 00:00:00

  • Optimal method of collection of first-void urine for diagnosis of Chlamydia trachomatis infection in men.

    abstract::First-void urine (FVU) is the preferred specimen for the diagnosis of urogenital Chlamydia trachomatis infection in men. We have developed FirstBurst, a urine collection device that collects the first 4 to 5 ml of FVU and yields a specimen with a sixfold higher C. trachomatis organism load than the regular urine cup b...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02241-07

    authors: Wisniewski CA,White JA,Michel CE,Mahilum-Tapay L,Magbanua JP,Nadala EC Jr,Barber PJ,Goh BT,Lee HH

    更新日期:2008-04-01 00:00:00

  • Confirmation of the distinct genotype groups within the form species Candida parapsilosis.

    abstract::The DNA relatedness and restriction fragment length polymorphism patterns of whole-cell DNA and the general phenotypic properties of 14 isolates of the form species Candida parapsilosis representing three diverse genetic groups were compared. The data confirm the unrelatedness of the three groups at the species level....

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.36.1.216-218.1998

    authors: Roy B,Meyer SA

    更新日期:1998-01-01 00:00:00

  • Enzyme immunoassay for detection of antibody to toxins A and B of Clostridium difficile.

    abstract::An enzyme immunoassay (enzyme-linked immunosorbent assay [ELISA]) to detect hamster antibody to toxins A and B of Clostridium difficile was developed in which toxin preparations are used to coat the solid phase. The specificity of the assay was supported by blocking tests with the toxin preparations and proteins from ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.18.2.242-247.1983

    authors: Viscidi RP,Yolken RH,Laughon BE,Bartlett JG

    更新日期:1983-08-01 00:00:00