Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.

Authors:
Address: Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, China.
Journal:

Publication:
Free Text: Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.

abstract

BACKGROUND:

Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel Loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain.

FINDINGS:

The assay was performed with Bst DNA polymerase at 65°C for 1 h. The detection limit of the 529 bp-LAMP assay was as low as 0.6 fg of T. gondii DNA. The sensitivity of this assay was 100 and 1000 fold higher than that of the LAMP targeting B1 gene (B1-LAMP) and nested PCR targeting 529 bp repeat element (529 bp-nested PCR), respectively. The specificity of the 529 bp-LAMP assay was determined using the DNA samples of Trypanosoma evansi, Plasmodium falciparum, Paragonimus westermani, Schistosoma japonicum, Fasciola hepatica and Angiostrongylus cantonensis. No cross-reactivity with the DNA of any parasites was found. The assay was able to detect T. gondii DNA in all mouse blood samples at one day post infection (dpi).

CONCLUSIONS:

We report the following findings: (i) The detection limit of the 529 bp-LAMP assay is 0.6 fg of T. gondii DNA; (ii) The assay does not involve any cross-reactivity with the DNA of other parasites; (iii) This is the first report on the application of the LAMP assay for early diagnosis of toxoplasmosis in blood samples from experimentally infected mice. Due to its simplicity, sensitivity and cost-effectiveness for common use, we suggest that this assay should be used as an early diagnostic tool for health control of toxoplasmosis.

Related Articles
Urine sample used for detection of toxoplasma gondii infection by loop-mediated isothermal amplification (LAMP).
Folia Parasitol (Praha). 2012
Urine sample used for detection of toxoplasma gondii infection by loop-mediated isothermal amplification (LAMP).
Hu X, Pan CW, Li YF, Wang H, Tan F. Folia Parasitol (Praha). 2012 Feb; 59(1):21-6.
Comparison of loop-mediated isothermal amplification (LAMP) and real-time PCR method targeting a 529-bp repeat element for diagnosis of toxoplasmosis.
Vet Parasitol. 2012
Comparison of loop-mediated isothermal amplification (LAMP) and real-time PCR method targeting a 529-bp repeat element for diagnosis of toxoplasmosis.
Lin Z, Zhang Y, Zhang H, Zhou Y, Cao J, Zhou J. Vet Parasitol. 2012 Apr 30; 185(2-4):296-300. Epub 2011 Oct 18.
Loop-mediated isothermal amplification: rapid detection of Angiostrongylus cantonensis infection in Pomacea canaliculata.
Parasit Vectors. 2011
Loop-mediated isothermal amplification: rapid detection of Angiostrongylus cantonensis infection in Pomacea canaliculata.
Chen R, Tong Q, Zhang Y, Lou D, Kong Q, Lv S, Zhuo M, Wen L, Lu S. Parasit Vectors. 2011 Oct 25; 4:204. Epub 2011 Oct 25.
Toxoplasma gondii: sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method.
Exp Parasitol. 2009
Toxoplasma gondii: sensitive and rapid detection of infection by loop-mediated isothermal amplification (LAMP) method.
Zhang H, Thekisoe OM, Aboge GO, Kyan H, Yamagishi J, Inoue N, Nishikawa Y, Zakimi S, Xuan X. Exp Parasitol. 2009 May; 122(1):47-50. Epub 2009 Feb 1.
Review [Problems and limitations of conventional and innovative methods for the diagnosis of Toxoplasmosis in humans and animals].
Parassitologia. 2004
Review [Problems and limitations of conventional and innovative methods for the diagnosis of Toxoplasmosis in humans and animals].
Piergili Fioretti D. Parassitologia. 2004 Jun; 46(1-2):177-81.

To top • Home

TraveldoctorOnline 2001 • Disclaimer • webmaster

The contents within traveldoctoronline are presented only for informational purposes and cannot substitute for professional health care or any other medical treatment.All users of this website with health problems should be oblige always to consult their medical doctor before starting any treatment.