Development of loop-mediated isothermal amplification (LAMP) as a diagnostic tool of toxoplasmosis

Krasteva, Donika; Toubiana, Mylène; Hartati, Sri; Kusumawati, Asmarani; Dubremetz, Jean‐François; Widada, Joannes Sri

doi:10.1016/j.vetpar.2009.03.014

Cited by 31 publications

(16 citation statements)

References 12 publications

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“…[23]. Thereafter, the LAMP assay was developed and evaluated for the detection of T. gondii infection from the lymph nodes of pigs [24], various organs harvested from mice [25], and blood samples from patients [26]. …”

Section: Resultsmentioning

confidence: 99%

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

Kong

Tong

et al. 2012

Parasites Vectors

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BackgroundToxoplasmosis 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.FindingsThe 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).ConclusionsWe 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.

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Section: Resultsmentioning

confidence: 99%

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

Kong

Tong

et al. 2012

Parasites Vectors

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“…The method uses four to six primers that recognize six to eight regions of the target DNA, which eliminates nonspecific binding, thereby ensuring the specificity of LAMP (30). Recently, this method was found to be a powerful diagnostic tool, and LAMP targeting the SAG1 gene (SAG1-LAMP) and the 200-to 300-fold repetitive 529-bp gene of T. gondii was successfully used for the diagnosis of toxoplasmosis in animal models, especially mice (24,37). The SAG1-LAMP was applied to detect the presence of T. gondii in infected mouse organs.…”

mentioning

confidence: 99%

“…The SAG1-LAMP was applied to detect the presence of T. gondii in infected mouse organs. This makes the method attractive for T. gondii detection in biopsy specimens (24). LAMP based on the 529-bp repetitive gene has also been shown to be useful for detection of T. gondii DNA extracted from lymph nodes of veterinary samples (37).…”

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confidence: 99%

Specific, Sensitive, and Rapid Diagnosis of Active Toxoplasmosis by a Loop-Mediated Isothermal Amplification Method Using Blood Samples from Patients

et al. 2010

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Loop-mediated isothermal amplification (LAMP), a rapid nucleic acid amplification method, was developed for the clinical diagnosis of toxoplasmosis. Three LAMP assays based on the SAG1, SAG2, and B1 genes of Toxoplasma gondii were developed. The sensitivities and specificities of the LAMP assays were evaluated by comparison with the results of conventional nested PCR. The LAMP assays were highly sensitive and had a detection limit of 0.1 tachyzoite, and no cross-reactivity with the DNA of other parasites was observed. Blood was collected from 105 individuals to test the LAMP assays: 40 patients with active toxoplasmosis, 40 negative controls, and 25 patients with other parasitic infections. The SAG2-based LAMP (SAG2-LAMP) had a greater sensitivity (87.5%) than the SAG1-LAMP (80%), B1-LAMP (80%), and nested PCR (62.5%). All the LAMP assays and nested PCR were 100% specific. This is the first report of a study which applied the LAMP method to diagnose toxoplasmosis from human blood samples. Due to its simplicity, sensitivity, and specificity, LAMP is suggested as an appropriate method for routine diagnosis of active toxoplasmosis in humans.

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“…Several reports were confirmed the higher sensitivity of LAMP for detection of T. gondii than conventional PCR targeting 529 bp fragment of T. gondii genome (Fallahi et al 2014;Kong et al 2012;Lin et al 2012;Zhang et al 2009) or other specific targeting of T. gondii DNA regions (Fallahi et al 2014;Hu et al 2012;Kong et al 2012;Krasteva et al 2009;Lau et al 2010;Lin et al 2012;Sotiriadou and Karanis 2008;Zhang et al 2009). Until now, LAMP method was used for detection of T. gondii in pig (Lili et al 2014;Lin et al 2012;Qu et al 2013;Wang et al 2013;Zhang et al 2009), sheep (Lin et al 2012), mice (Hu et al 2012;Kong et al 2012;Krasteva et al 2009), and human (Fallahi et al 2014;Lau et al 2010). LAMP was also used for detection of T. gondii oocysts in water samples (GallasLindemann et al 2013;Koloren and Demirel 2013;Sotiriadou and Karanis 2008).…”

Section: Discussionmentioning

confidence: 82%

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

et al. 2015

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Toxoplasma gondii is one of the most common zoonotic parasitic diseases in human and warm-blooded animals worldwide. Birds are one of important intermediate hosts of T. gondii. The aim of this study is molecular detection of T. gondii in the house sparrow by LAMP and PCR methods in Tehran, Iran. A total 200 sparrows were captured in different regions of Tehran. DNA was extracted from tissue samples of each sparrow. LAMP and conventional PCR assays were carried out with a set of primers to detect the 529 bp fragment of T. gondii. LAMP and PCR were detected T. gondii from 17 (8.5 %) and 15 (7.5 %) of 200 sparrows respectively. These results indicated that sensitivity of LAMP was higher than conventional PCR. In our knowledge, this study is the first report of detection of T. gondii by LAMP method in bird hosts. Also, these findings provided an insight into epidemiological pattern of T. gondii infection in sparrow in Iran.

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Development of loop-mediated isothermal amplification (LAMP) as a diagnostic tool of toxoplasmosis

Cited by 31 publications

References 12 publications

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

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

Specific, Sensitive, and Rapid Diagnosis of Active Toxoplasmosis by a Loop-Mediated Isothermal Amplification Method Using Blood Samples from Patients

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

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