Simultaneous detection of Theileria annulata and Theileria orientalis infections using recombinase polymerase amplification

Hassan, Mohammad Farooque; Liu, Junlong; Sajid, Muhammad Sohail; Rashid, Imran; Mahmood, Altaf; Abbas, Qamar; Guan, Guiquan; Yin, Hong; Luo, Jianxun

doi:10.1016/j.ttbdis.2018.03.028

Cited by 15 publications

(7 citation statements)

References 17 publications

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“…Despite the diagnostic artifacts (low sensitivity and specificity: unable to detect Theileria species in carrier state and low parasitemia), the examination of piroplasms in Giemsa-stained blood smears is the most frequently used diagnostic technique in veterinary clinical settings to identify acute TT in disease-endemic regions of Pakistan. This is due to the unavailability of molecular and serological diagnostics and resource limitations in most rural veterinary hospitals and clinical settings [ 49 ]. The relative sensitivity and specificity of the blood microscopy, as a diagnostic technique, validated previous findings [ 38 , 45 ].…”

Section: Discussionmentioning

confidence: 99%

Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan

et al. 2021

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Theileria annulata is a tick-borne hemoprotozoan parasite responsible for tropical theileriosis in the bovine population, which causes substantial economic losses to the livestock sector. The present study has investigated, characterized, and shaped epidemiologic and phylogenetic profiles of T. annulata infection in the cattle population of central Khyber Pakhtunkhwa, Pakistan. A total of 600 blood samples were collected from cattle. Microscopy and PCR (18S rRNA taxonomic marker) assays were performed to detect T. annulata infection in cattle from the study area. The overall relative prevalence rates of T. annulata in the examined cattle population were 12.8% (microscopy) and 23.7% (PCR). District-wise analysis (microscopy/PCR) showed that cattle from district Mardan were found more infected (16.0%/28.0%), as compared to cattle from district Charsadda (13.5%/25.5%) and district Peshawar (9.0%/17.5%). Based on host demographic and ecological parameters analysis, theileriosis was found to be higher in young, female, crossbred, freely grazing, tick-infested, and irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, tick infestation, acaricides use, and feeding method were significant risk factors (P<0.05) whereas multivariate analysis indicated that host age, gender, tick infestation, acaricidal application, and feeding method were potential risk factors (P<0.05) for tropical theileriosis in the cattle population. Phylogenetic and sequence analysis showed that T. annulata 18S rRNA isolates shared homology and phylogeny with other isolates from Asia and Europe. This study has addressed the epidemiology and phylogeny of T. annulata circulating in bovid in the study area where gaps were still present. These findings will serve as a baseline and will facilitate future large-scale epidemiological investigations on tropical theileriosis in the cattle population at a national level.

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Section: Discussionmentioning

confidence: 99%

Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan

et al. 2021

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“…Many studies have shown that RPA can be performed at 25–45 °C without the use of complicated equipment, which overcomes environmental constraints. These advantages have allowed RPA to be used successfully in the detection of various pathogens such as viruses (Hou et al 2017 ; Tu et al 2017 ; Yang et al 2017 ; Miao et al 2019 ), bacteria (Gao et al 2018 ; Zhang et al 2019 ), and parasites (Castellanos-Gonzalez et al 2018 ; Hassan et al 2018 ; Wu et al 2019 ). Although RPA has been frequently applied to detect pathogens related to humans and domestic animals, its application in plant pathogen detection remains poorly studied.…”

Section: Introductionmentioning

confidence: 99%

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Song

et al. 2021

Phytopathol Res

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Phytophthora cactorum is a devastating pathogen that infects a wide range of plants and causes Phytophthora rot disease, which has resulted in great economic losses in crop production. Therefore, the rapid and practicable detection of P. cactorum is important for disease monitoring and forecasting. In this study, we developed a lateral flow recombinase polymerase amplification (LF-RPA) assay for the sensitive visual detection of P. cactorum. Specific primers for P. cactorum were designed based on the ras-related protein gene Ypt1; all 10 P. cactorum isolates yielded positive detection results, whereas no cross-reaction occurred in related oomycete or fungal species. The detection limit for the LF-RPA assay was 100 fg of genomic DNA under optimized conditions. Combined with a simplified alkaline lysis method for plant DNA extraction, the LF-RPA assay successfully detected P. cactorum in naturally diseased strawberry samples without specialized equipment within 40 min. Thus, the LF-RPA assay developed in this study is a rapid, simple, and accurate method for the detection of P. cactorum, with the potential for further application in resource-limited laboratories.

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“…Gebrekidan et al [ 20 ] reported the presence of T. orientalis in Pakistan for the first time from imported and native animals, reporting no T. orientalis in native animals by conventional PCR and low prevalence (6%) in native cattle as compared with imported animals (25.4%) using multiplexed tandem PCR. Recently, Hassan et al [ 21 , 22 ] determined the presence of Theileria spp. by using recombinase polymerase amplification.…”

Section: Introductionmentioning

confidence: 99%

Molecular survey of piroplasm species from selected areas of China and Pakistan

Hassan¹,

Liu

Rashid

et al. 2018

Parasites Vectors

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BackgroundPiroplasmosis is an important animal disease that is a major constraint to the development of the livestock industry, often resulting in significant economic losses. Therefore, there is an urgent need to further understand the etiology of this and other tick-borne infections.MethodsBlood samples were collected from tick-infested animals from the Chakwal, Jhang, and Faisalabad districts of Punjab, Pakistan and from peri-urban areas around Hohhot, Inner Mongolia, China to investigate the presence of Babesia and Theileria species. In total, 450 blood samples were collected with FTA cards from cattle of the study areas of Pakistan; the genomic (g)DNA of one hundred and twenty samples from cattle in Inner Mongolia were provided by the Lanzhou Veterinary Research Institute, China. Following the extraction of gDNA, the 18S rRNA gene (V4 hypervariable region) of piroplasms was amplified in all samples using semi-nested PCR. Positively identified samples were sequenced for the identification of Theileria and Babesia species. The partial full-length sequence of 18S rDNA was amplified for species confirmation of Theileria-positive samples, whereas the RAP-1c gene was amplified for Babesia bigemina-positive samples.ResultsSemi-nested PCR results revealed that 144 (25.26%) samples were positive for piroplasms. Theileria annulata was the most prevalent species (115/144; 20.17%), followed by Theileria orientalis (16/144; 2.80%). Among Babesia, the only species recorded was Babesia bigemina (13/144; 2.28%).ConclusionThe present study reveals new data on the prevalence of piroplasm species in bovine populations of selected areas of China and Pakistan and their phylogenetic relationships. It is also the first detailed report of T. orientalis from native animals in Pakistan.

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Simultaneous detection of Theileria annulata and Theileria orientalis infections using recombinase polymerase amplification

Cited by 15 publications

References 17 publications

Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan

Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan

Rapid and simple detection of Phytophthora cactorum in strawberry using a coupled recombinase polymerase amplification–lateral flow strip assay

Molecular survey of piroplasm species from selected areas of China and Pakistan

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