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Table 2 Evaluation results of LAMP and PCR specificity for the detection of Toxoplasma B1 gene

From: Rapid diagnosis of Toxoplasma gondii using loop-mediated isothermal amplification assay in camels and small ruminants

DNA extraction PCR LAMP
Toxoplasma B1 gene Specificity Toxoplasma B1 gene Specificity
E. coli 100% 100%
Salmonella species   
Giardia lamblia   
N. caninum   
Tissue Oocyte Toxoplasma RH +   +  
Toxoplasma RH (tachyzoites) mice +   +  
Toxoplasma (tachyzoites) bovine +   +  
Toxoplasma (tachyzoites) small ruminants +   +  
Toxoplasma (tachyzoites) equines +   +  
Toxoplasma RH (tachyzoites) SPF +   +  
Negative control (dH2O)   
Negative control SPF with PBS pH 7.2   
Negative control proteineal fluid from mice inoculated with PBS   
  1. Table 2 shows that the designed four LAMP primers were used to amplify the AF and or RE fragment of extracted DNA from pathogens, including E. coli, Staphylococcus spp, Salmonella species, Giardia lamblia, N. caninum, Tissue cyst   Toxoplasma RH, Toxoplasma RH (tachyzoites) mice, Toxoplasma (tachyzoites) bovine and negative control SPF. with PBS pH 7.2, negative control proteineal fluid from mice inoculated with PBS as the template, no such target was observed. It is suggested that the LAMP assay with target-specific primers were showed high specificity in the detection of T. gondii. LAMP assay sensitivity: The LAMP assay's sensitivity was tested by detecting DNA amplification with various copies, ranging from 101 to 106. The sensitivity of LAMP was also tested using genomic DNA isolated from various quantities of tachyzoites as templates. When genomic DNA from at least five tachyzoites was used, the amplified DNA fragments were visible. Although there was no DNA product in the condition with only one tachyzoite's genomic DNA, there was no visible change in the reactions of the amplified target segment. According to the results, the LAMP assay was very sensitive in detecting Toxoplasma even with only 10 copies of the target fragment or five tachyzoites in the sample