Korean J Parasitol.  2013 Apr;51(2):237-241. 10.3347/kjp.2013.51.2.237.

Sensitive and Rapid Detection of Giardia lamblia Infection in Pet Dogs using Loop-Mediated Isothermal Amplification

Affiliations
  • 1College of Veterinary Medicine, South China Agricultural University, 483 Wushan Street, Tianhe District, Guangzhou, Guangdong Province 510642, People's Republic of China. gqli@scau.edu.cn

Abstract

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10(-1) to 10(-5) ng/microl for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63degrees C by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1alpha) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

Keyword

Giardia lamblia; loop-mediated isothermal amplification (LAMP); PCR; EF1alpha gene; dog

MeSH Terms

Animals
Dog Diseases/*diagnosis/parasitology
Dogs
Feces/parasitology
Giardia lamblia/genetics/*isolation & purification
Giardiasis/diagnosis/parasitology/*veterinary
Humans
Molecular Diagnostic Techniques/*methods
Molecular Sequence Data
Nucleic Acid Amplification Techniques/*methods
Pets
Sensitivity and Specificity
Sequence Analysis, DNA
Temperature
Time Factors
Veterinary Medicine/*methods
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