A polymerase chain reaction (PCR) using heat-stable Taq polymerase is described for the specific detection of Mycobacterium leprae, the causative agent of leprosy. A set of primers was selected on the basis of the nucleotide sequence of a gene encoding the 36 kDa antigen of M. leprae. With this set of primers in the PCR, M. leprae could be detected specifically with a detection limit approximating one bacterium. This PCR appears to meet the criteria of specificity and sensitivity required for a useful tool in epidemiology and eventually for the control of leprosy.
Clark-CurtissJ.E.1988; Molecular relatedness among Mycobacterium leprae isolates defined using recombinant DNA probes (Abstracts of the 13th International Leprosy Congress ’88).. Health Cooperation Papers 9:125
Clark-CurtissJ.E.,
JacobsW.R.,
DochertyM.A.,
RitchieL.R.,
CurtissR.III1985; Molecular analysis of DNA and construction of genomic libraries of Mycobacterium leprae.. Journal of Bacteriology 161:1093–1102
EisenachK.D.,
CrawfordJ.T.,
BatesJ.H.1988; Repetitive DNA sequences as probes for Mycobacterium tuberculosis.. Journal of Clinical Microbiology 26:2240–2245
Van EysG.J.J.M.,
ZaalJ.,
SchooneG.J.,
TerpstraW.J.1988; DNA hybridization with hardjobovis-specific recombinant probes as a method for type discrimination of Leptospirainterrogans serovar hardjo.. Journal of General Microbiology 134:567–574
FineP.E.M.,
PonnighausJ.M.,
BurgessP.,
ClarksonJ.A.,
DraperC.C.1988; Seroepide- miological studies of leprosy in Northern Malawi based on an enzyme-linked immunosorbent assay using synthetic glycoconjugate antigen.. International Journal of Leprosy 56:243–254
HartskeerlR.A.,
SuykerbuykM.E.G.,
GuptaR.,
TholeJ.E.R.1988; Selection of recombinant DNA clones expressing the 12K and 36K M. leprae antigens in E. coli (Abstracts of the 13th International Leprosy Congress ’88).. Health Cooperation Papers 9:129
KirchheimerW.F.,
StorrsE.E.1971; Attempts to establish the armadillo (Dasypus novemcinctusLinn.) as a model for the study of leprosy.. International Journal of Leprosy 39:693–703
KoganS.C.,
DohertyM.,
GitschierJ.1987; An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A.. New England Journal of Medicine 317:985–990
KolkA.H.J.,
HoM.L.,
KlatserP.R.,
EggelteT.A.,
PortaelsF.1985; Production of monoclonal antibodies against Mycobacterium lepraeand Armadillo-derived mycobacteria.. Annales de I’Institut Pasteur 136B:217–221
MehraV.,
SweetserD.,
YoungR.A.1986; Efficient mapping of protein antigenic determinants.. Proceedings of the National Academy of Sciences of the United States of America 83:7013–7017
OuC.,
KwokS.,
MitchellS.W.,
MackD.H.,
SninskyJ.J.,
KrebsJ.W.,
FeorinoP.,
WarfieldD.,
SchochetmanG.1988; DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells.. Science 239:295–297
PortaelsF.,
PattynS.R.1982; Isolation of fastidiously growing mycobacteria from armadillo livers infected with Mycobacterium leprae.. International Journal of Leprosy 50:370–374
SaikiR.K.,
ScharfS.,
FaloonaF.,
MullisK.B.,
HornG.T.,
ErlichH.A.,
ArnheimN.1985; Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.. Science 230:1350–1354
SaikiR.K.,
GelfandD.H.,
StoffelS.,
ScharfS.J.,
HiguchiR.,
HornG.T.,
MullisK.B.,
EhrlichH.A.1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.. Science 239:487–491
ShibataD.K.,
ArnheimN.,
MartinW.J.1988; Detection of human papilloma virus in paraffin- embedded tissue using the polymerase chain reaction.. Journal of Experimental Medicine 167:225–230
ShinnickT.M.,
SweetserD.,
TholeJ.,
VanembdenJ.,
YoungR.A.1987; The etiological agents of leprosy and tuberculosis share an immuno-reactive protein antigen with the vaccine strain Mycobacterium bovis BCG.. Infection and Immunity 55:1932–1935
ShinnickT.M.,
VodkinM.H.,
WilliamsJ.C.1988; The Mycobacterium tuberculosis 65-kilodalton antigen is a heat shock protein which corresponds to common antigen and to the Escherichia coli GroEL protein.. Infection and Immunity 56:446–451
ShoemakerS.A.,
FisherJ.H.,
ScogginH.1985; Techniques of DNA hybridization detect small numbers of mycobacteria with no cross-hybridization with nonmycobacterial respiratory organisms.. Annual Review of Respiratory Diseases 131:760–763
TholeJ.E.R.1988A major immunogenic mycobacterial antigen: its molecular characterization and its implication in adjuvant arthritis. PhD Thesis University of Amsterdam.:
TholeJ.E.R.,
KeulenW.C.,
KolkA.H.J.,
GroothuisD.G.,
BerwaldL.G.,
TietsjemaR.H.,
Ven EmbdenJ.D.A.1987; Characterization, sequence determination, and immunogenicity of a 64-kilodalton protein of Mycobacterium bovisBCG expressed in Escherichia coli.. Infection and Immunity 50:800–806
TholeJ.E.R.,
HinderssonP.,
De BruynJ.,
CremersF.,
Van Der ZeeJ.,
De CockH.,
TommassenJ.,
Van EdenW.,
Van EmbdenJ.D.A.1988; Antigenic relatedness of a strongly immunogenic 65 kDa mycobacterial protein antigen with a similar sized ubiquitous bacterial common antigen.. Microbial Pathogenesis 4:71–83
De WitM.Y.L.,
KlatserP.R.1988; Purification and characterization of a 36 kDa antigen of Mycobacterium leprae.. Journal of General Microbiology 134:1541–1548
WongC.,
DowlingC.E.,
SaikiR.K.,
HiguchiR.G.,
ErlichH.A.,
KazazianH.H.1987; Characterization of β-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA.. Nature; London: 330:384–386
World Health Organization1980; Purification of M. leprae. Report of the 5th meeting of the Scientific Working Group on the Immunology of Leprosy (IMMLEP). TDR/IMMLEP-SWG(5)/80.3.. Geneva:: World Health Organization.;
World Health Organization1988; WHO Expert Committee on Leprosy: 6th Report. Technical Report Series, no. 768.. Geneva:: World Health Organization.;
YaegerH.JrLacyJ.,
SmithL.R.,
LemaisterC.A.1966; Quantitative studies of mycobacterial populations in sputum and saliva.. American Review of Respiratory Diseases 95:998–1004