کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5545601 1555633 2017 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Research paperRecombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم دامی و جانورشناسی
پیش نمایش صفحه اول مقاله
Research paperRecombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment
چکیده انگلیسی


- Develop a recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water.
- The amplification product was visualized by lateral flow (LF) strip in 5 min.
- The sensitivity of RPA was 10 times higher than that of nest PCR.
- B1-LF-RPA a promising method as a molecular detection tool for T. gondii.

Toxoplasma gondii infects all warm-blooded vertebrates, resulting in a great threat to human health and significant economic loss to the livestock industry. Ingestion of infectious oocysts of T. gondii from the environment is the major source of transmission. Detection of T. gondii oocysts by existing methods is laborious, time-consuming and expensive. The objective of the present study was to develop a recombinase polymerase amplification (RPA) method combined with a lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water. The DNA of T. gondii oocysts was amplified by a pair of specific primers based on the T. gondii B1 gene over 15 min at a constant temperature ranging from 30 °C to 45 °C using RPA. The amplification product was visualized by the lateral flow (LF) strip within 5 min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species. Fifty environmental samples were further assessed for the detection validity of the LF-RPA assay (B1-LF-RPA) and compared with nested PCR based on the B1 gene sequence. The B1-LF-RPA and nested PCR both showed that 5 out of the 50 environmental samples were positive. The B1-LF-RPA method was also proven to be sufficiently tolerant of existing inhibitors in the environment. In addition, the advantages of simple operation, speediness and cost-effectiveness make B1-LF-RPA a promising molecular detection tool for T. gondii.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Veterinary Parasitology - Volume 243, 30 August 2017, Pages 199-203
نویسندگان
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