Polymerase Spiral Reaction (PSR) as a novel rapid colorimetric isothermal point of care assay for detection of Trypanosoma evansi genomic DNA

Sharma, Diksha; Gupta, S. C.; Sethi, Khushboo; Kumar, Sanjay; Kumar, Rajender

doi:10.1016/j.vetpar.2021.109644

Cited by 7 publications

(6 citation statements)

References 23 publications

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“…Some will be easy to develop since all the parameters, methods and reagents are available, such as a pan- Trypanosoma ELISA or POCD based on in vitro - produced parasites. Some will require short- or medium-term research plans, such as RNA-based methods or PSR (or even SHERLOCK), for which preliminary supporting work has recently been published [ 37 , 153 ]. Others may require long-term research before field implementation, such as a card agglutination test for IgM detection targeting T. vivax , T. equiperdum or T. lewisi , provided a dominant early antigen is identified, or a species-specific POCD based on recombinant antigens.…”

Section: Discussionmentioning

confidence: 99%

“…Point-of-care diagnostics based on DNA amplification, such as LAMP [ 152 ], have been developed, but have not been successfully applied in the field. New developments, such as RPA-LFD or even PSR [ 37 ] are promising. Recent studies have demonstrated a high amount of small trypanosome RNA in the serum of cattle with active infections [ 39 , 153 ].…”

Section: Future Prospectsmentioning

confidence: 99%

“…Although they were claimed to be efficient and applicable in the field [34][35][36], this has never really been the case. The new polymerase spiral reaction (PSR) method [37] may be suitable for field diagnosis in real time, but it still requires comprehensive field validation. Finally, the new and promising spliced-leader RNA (SL-RNA) detection method is applied to a short and conserved RNA sequence linked to the 5' end of each trypanosome pre-messenger RNA (mRNA) [38].…”

Section: Detection Of An "Active Infection"mentioning

confidence: 99%

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Diagnosis of animal trypanosomoses: proper use of current tools and future prospects

et al. 2022

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Reliable diagnostic tools are needed to choose the appropriate treatment and proper control measures for animal trypanosomoses, some of which are pathogenic. Trypanosoma cruzi, for example, is responsible for Chagas disease in Latin America. Similarly, pathogenic animal trypanosomoses of African origin (ATAO), including a variety of Trypanosoma species and subspecies, are currently found in Africa, Latin America and Asia. ATAO limit global livestock productivity and impact food security and the welfare of domestic animals. This review focusses on implementing previously reviewed diagnostic methods, in a complex epizootiological scenario, by critically assessing diagnostic results at the individual or herd level. In most cases, a single diagnostic method applied at a given time does not unequivocally identify the various parasitological and disease statuses of a host. These include “non-infected”, “asymptomatic carrier”, “sick infected”, “cured/not cured” and/or “multi-infected”. The diversity of hosts affected by these animal trypanosomoses and their vectors (or other routes of transmission) is such that integrative, diachronic approaches are needed that combine: (i) parasite detection, (ii) DNA, RNA or antigen detection and (iii) antibody detection, along with epizootiological information. The specificity of antibody detection tests is restricted to the genus or subgenus due to cross-reactivity with other Trypanosoma spp. and Trypanosomatidae, but sensitivity is high. The DNA-based methods implemented over the last three decades have yielded higher specificity and sensitivity for active infection detection in hosts and vectors. However, no single diagnostic method can detect all active infections and/or trypanosome species or subspecies. The proposed integrative approach will improve the prevention, surveillance and monitoring of animal trypanosomoses with the available diagnostic tools. However, further developments are required to address specific gaps in diagnostic methods and the sustainable control or elimination of these diseases. Graphical Abstract

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Section: Discussionmentioning

confidence: 99%

Section: Future Prospectsmentioning

confidence: 99%

Section: Detection Of An "Active Infection"mentioning

confidence: 99%

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Diagnosis of animal trypanosomoses: proper use of current tools and future prospects

et al. 2022

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“…PSR assay for clinical diagnosis of viral and bacterial diseases and food pathogens are highly specific to their respective target nucleic acid sequences ( Liu et al, 2015 ; Gupta et al, 2017 ; Malla et al, 2018 ; Ji et al, 2019 ; Milton et al, 2020 , 2021 ; Sharma et al, 2020 , 2022 ; Tomar et al, 2020 ; Maiti et al, 2022 ). The PSR assay reported in the present study is specific to T. palmi mtCOIII region and does not cross-react to predominant thrips species.…”

Section: Discussionmentioning

confidence: 99%

“…It does not require any sophisticated equipment, and reagents are readily available. PSR has been successfully implemented in clinical settings and food testing ( Liu et al, 2015 ; Gupta et al, 2017 ; Malla et al, 2018 ; Ji et al, 2019 ; Milton et al, 2020 ; Sharma et al, 2020 , 2022 ; Tomar et al, 2020 ; Maiti et al, 2022 ). This is the first experimental demonstration where PSR has been utilized for the detection of insect species.…”

Section: Introductionmentioning

confidence: 99%

Development of a Polymerase Spiral Reaction-Based Isothermal Assay for Rapid Identification of Thrips palmi

Jangra

Ghosh

Mukherjee

et al. 2022

Front. Mol. Biosci.

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Thrips cause considerable economic losses to a wide range of food, feed, and forest crops. They also transmit several plant viruses. Being cryptic, it is often difficult to distinguish thrips species in crops and large consignments by conventional methods. Melon thrips (Thrips palmi Karny, Thysanoptera: Thripidae) is an invasive insect pest of vegetables, legumes, and ornamentals besides being vector to several viruses. It poses a threat to domestic and international plant biosecurity and can invade and establish in new areas. Here, we report a polymerase spiral reaction (PSR)-based isothermal assay for rapid, sensitive, specific, low-cost, and on-site detection of T. palmi. To the best of our knowledge, this is the first application of PSR in the identification of any insect species. A primer pair designed based on 3′-polymorphism of mtCOIII region can specifically identify T. palmi without any cross-reactivity with predominant thrips species. The assay uses crude lysate of a single thrips saving time and reagents involved in nucleic acid extraction. The presence of T. palmi is visualized by the appearance of bright fluorescence under ultraviolet light or a change in reaction color thus avoiding gel electrophoresis steps. The entire process can be completed in 70 min on-site using only an ordinary water bath. The assay is sensitive to detecting as little as 50 attograms of T. palmi template. The assay was validated with known thrips specimens and found to be efficient in diagnosing T. palmi under natural conditions. The described method will be useful for non-expert personnel to detect an early infestation, accidental introduction to a new area, restrict the spread of diseases and formulate appropriate management strategies.

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An overview of diagnostic evaluation of parasitic diseases of dogs and cats

Gupta¹,

Harit²,

Sharma³

et al. 2023

Organ-Specific Parasitic Diseases of Dogs and Cats

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Polymerase Spiral Reaction (PSR) as a novel rapid colorimetric isothermal point of care assay for detection of Trypanosoma evansi genomic DNA

Cited by 7 publications

References 23 publications

Diagnosis of animal trypanosomoses: proper use of current tools and future prospects

Diagnosis of animal trypanosomoses: proper use of current tools and future prospects

Development of a Polymerase Spiral Reaction-Based Isothermal Assay for Rapid Identification of Thrips palmi

An overview of diagnostic evaluation of parasitic diseases of dogs and cats

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