Detection of Acute Toxoplasmosis in Pigs Using Loop-Mediated Isothermal Amplification and Quantitative PCR
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A loop-mediated isothermal amplification (LAMP) assay allows rapid diagnosis of Toxoplasma gondii infection. In the present study, the LAMP assay was evaluated using blood from both naturally and experimentally infected pigs. The sensitivity of the LAMP assay was compared with that of Q-PCR. Both assays detected T. gondii in the blood of experimentally infected pigs, with 100% agreement. In infected blood samples, the parasite was detected as early as 2 days post-infection and reached a peak in 3-5 days. In 216 field serum samples, the detection rates of LAMP and Q-PCR assays were 6.9% and 7.8%, respectively. This result indicates that the sensitivity of the LAMP assay was slightly lower than that of the Q-PCR assay. However, the LAMP may be an attractive diagnostic method in conditions where sophisticated and expensive equipment is unavailable. This assay could be a powerful supplement to current diagnostic methods.
Abstract
A loop-mediated isothermal amplification (LAMP) assay allows rapid diagnosis of Toxoplasma gondii infection. In the present study, the LAMP assay was evaluated using blood from both naturally and experimentally infected pigs. The sensitivity of the LAMP assay was compared with that of Q-PCR. Both assays detected T. gondii in the blood of experimentally infected pigs, with 100% agreement. In infected blood samples, the parasite was detected as early as 2 days post-infection and reached a peak in 3-5 days. In 216 field serum samples, the detection rates of LAMP and Q-PCR assays were 6.9% and 7.8%, respectively. This result indicates that the sensitivity of the LAMP assay was slightly lower than that of the Q-PCR assay. However, the LAMP may be an attractive diagnostic method in conditions where sophisticated and expensive equipment is unavailable. This assay could be a powerful supplement to current diagnostic methods.
참고문헌 (21)
- Nielsen HV, "Complete protection against lethal Toxoplasma gondii infection in mice immunized with a plasmid encoding the SAG1 gene", Infect Immun, 67, pp.6358~6363 (1999)
- Dubey JP, "Persistence of immunity to toxoplasmosis in pigs vaccinated with a non-persistent strain of Toxoplasma gondii", Am J Vet Res, 55, pp.982~987 (1994)
- Foulon W, "Prenatal diagnosis of congenital toxoplasmosis : a multicenter evaluation of different diagnostic parameters", Am J Obstet, 181, pp.843~847 (1999)
- Pelloux H, "A new set of primers for the detection of Toxoplasma gondii in amniotic fluid using polymerase chain reaction", FEMS Microbiol Lett, 138, pp.11~15 (1996)
- Wang YH, "Development of an immunochromatographic strip for the rapid detection of Toxoplasma gondii circulating antigens", Parasitol Int, 60, pp.105~107 (2011)
- Zhang HS, "Toxoplasma gondii : Sensitive and rapid detection of infection by loop-mediated isothermal amplification(LAMP)method", Exp Parasitol, 122, pp.47~50 (2009)
- Switaj K, "Recent trends in molecular diagnostics for Toxoplasma gondii infections", Clin Microbiol Infect, 11, pp.170~176 (2005)
- Notomi T, "Loop-mediated isothermal amplification of DNA", Nucleic Acids Res, E28, pp.I~VII (2000)
- Mori Y, "Detection of loop-mediated isothermal reaction by turbidity derived from magnesium pyrophosphate formation", Biochem Biophys Res Commun, 289, pp.150~154 (2001)
- Tomita N, "Loop-mediated isothermal amplification(LAMP)of gene sequences and simple visual detection of products", Nat Protoc, 3, pp.877~882 (2008)
- Qiao YM, "Loop-mediated isothermal amplification for rapid detection of Bacillus anthracis spores", Biotech Lett, 29, pp.1939~1946 (2007)
- Kuboki N, "Loop-mediated isothermal amplification for detection of African trypanosomes", J Clin Microbiol, 41, pp.5517~5524 (2003)
- Poon LLM, "Sensitive and inexpensive molecular test for Falciparum Malaria : detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification", Clin Chem, 52, pp.303~306 (2006)
- Thekisoe OMM, "Stability of loop-mediated isothermal amplification(LAMP)reagents and its amplification efficiency on crude trypanosome DNA templates", J Vet Med Sci, 71, pp.471~475 (2009)
- Kong QM, "Loop-mediated isothermal amplification(LAMP) : Early detection of Toxoplasma gondii infection in mice", Parasit Vectors, 5, pp.2~ (2012)
- Krasteva D, "Development of loop-mediated isothermal amplification(LAMP)as a diagnostic tool of toxoplasmosis", Vet Parasitol, 162, pp.327~331 (2009)
- Lau YL, "Specific, sensitive, and rapid diagnosis of active toxoplasmosis by a loop-mediated isothermal amplification method using blood samples from patients", J Clin Microbiol, 48, pp.3698~3702 (2010)
- Lin ZB, "Comparison of loop-mediated isothermal amplification(LAMP)and real-time PCR method targeting a 529-bp repeat element for diagnosis of toxoplasmosis", Vet Parasitol, 185, pp.296~300 (2012)
- Sotiriadoua I, "Evaluation of loop-mediated isothermal amplification for detection of Toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test(IFT)", Diagn Microbiol Infect Dis, 62, pp.357~365 (2008)
- Wang YH, "The study of antigenic epitopes from Toxoplasma gondii excreted", secreted antigens and surface protein 1, 2011, 19~20
- Kim SK, "Stage specific expression of surface antigens by Toxoplasma gondii as a mechanism to facilitate parasite persistence", J Immunol, 174, pp.8038~8048 (2005)