RPB0569

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Toxoplasma gondii Toxoplasma gondii 5811 Eucoccidiorida Sarcocystidae Toxoplasma Toxoplasma gondii Eukaryota

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM) GC Content(%) Predicted Melting Temperature(℃) Molecular Weight(g/moles) Positions in GenBank accession number
mF1 AGCCAGAGAAGGGACAGAAGTCGAAGGGGACT 32 \ 56.25 68.86 10030.58 \
mR3 CAGGAAAAGCAGCGAAGCTGGAAACATCTT 30 \ 46.67 63.02 9282.12 \
529-Pe AGCCGTCTTGGAGGAGAGATATCAGGACTG(FAM-dT)AG(THF)A(BHQ1-dT)GAAGGCGAGGGTGAG-P 48 \ 56.25 73.26 15100.83 \

Gene Description

Target Gene GenBank ID
529 gene AF146527.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
The RT-RAA assay established in this study, with the advantages of reliability, rapidity, and high sensitivity, is a reasonable alternative to RT-qPCR for point-of-care detection. RT-RAA Oligo 7 software 25 min 36 °C \ 10^2copies of per reaction \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2021 Development of a real-time recombinase-aided amplification (RT-RAA) molecular diagnosis assay for sensitive and rapid detection of Toxoplasma gondii Zhao-Hua Wang,Wei Zhang,Xiu-Zhong Zhang,Xin-Ran Yao,Wei Huang,Hong Jia,Xiao-Lei Liu,Shao-Hua Hou,Xiao-Jia Wang Veterinary parasitology 34384956 10.1016/j.vetpar.2021.109489

Development of a real-time recombinase-aided amplification (RT-RAA) molecular diagnosis assay for sensitive and rapid detection of Toxoplasma gondii

Author(s):

Zhao-Hua Wang,Wei Zhang,Xiu-Zhong Zhang,Xin-Ran Yao,Wei Huang,Hong Jia,Xiao-Lei Liu,Shao-Hua Hou,Xiao-Jia Wang

Journal:

Veterinary parasitology

Year:

2021

Abstract:

Toxoplasma gondii, a protozoan intracellular parasite, is present in a wide range of hosts, including virtually all species of warm-blooded vertebrates. Toxoplasmosis spreads to humans through a variety of pathways, including contaminated food or water, and close contact with various types of domestic animals. It poses a severe threat to human health, and contributes to important economic losses, not only in cost-of-illness but also in surveillance programs. It is thus necessary to develop a rapid point-of-care field diagnostic technology to control or prevent pathogen transmission to economically important livestock animals, domestic animals, and human beings. In this study, we develop a real-time isothermal amplification method capable of detecting the T. gondii genome in swine and feline blood samples. This method can detect toxoplasma genome with a lowest detection limit of 102 copies of per reaction under optimal reaction conditions of 36 °C for 25 min. The assay displayed advantages in sensitivity and specificity in comparison to traditional real-time PCR, and can be performed in a portable instrument.