Sensitive, specific, and rapid detection of Leishmania donovani DNA by loop-mediated isothermal amplification.
We have applied a loop-mediated isothermal amplification (LAMP) technique to detect Leishmania donovani DNA. The LAMP technique detected 1 fg of L. donovani DNA, which was 10-fold more sensitive than a conventional polymerase chain reaction (PCR). All nested PCR-positive blood samples from visceral leishmaniasis patients were positive with the LAMP technique, and DNA samples from L. infantum, L. major, L. mexicana, L. tropica, L. braziliensis, Plasmodium falciparum, and healthy humans were negative with the LAMP technique. The advantages of the LAMP method are its shorter reaction time, a lack of requirement of sophisticated equipment, and visual judgment of positivity based on the turbidity of reaction mixture. Our LAMP technique can be a better alternative to a conventional PCR, especially under field conditions.
Takagi
Hidekazu
H
Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
Itoh
Makoto
M
Islam
Mohammad Zahidul
MZ
Razzaque
Abdur
A
Ekram
A R M Saifuddin
AR
Hashighuchi
Yoshihisa
Y
Noiri
Eisei
E
Kimura
Eisaku
E
D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics*; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
D016054:DNA, Protozoan