Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus

Shalaby, Mohamed A.; El‐Deeb, Ayman H.; El‐Tholoth, Mohamed; Hoffmann, Donata; Czerny, Claus-Peter; Hufert, Frank T.; Weidmann, Manfred; Wahed, Ahmed Abd El

doi:10.1186/s12917-016-0875-5

Cited by 37 publications

(20 citation statements)

References 33 publications

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“…Lumpy skin disease (LSD) mainly infects cattle and is characterized by fever, lymphadenitis, edema of subcutaneous cellular tissue and viscera, cutaneous nodules (lumps), ocular discharge, and inflammation of the mucosae (Prozesky and Barnard 1982 ). It is a transmissible disease that is transferred by various arthropods (Chihota et al 2003 ) and causes significant economic losses because of cattle exhaustion, hide damage, infertility, mastitis, and reduced milk production and up to 20% mortality is reported (Shalaby et al 2016 ). LSD has been reported in other domestic species and wildlife naturally and through experimental infection (Young et al 1970 ; Usadov et al 2018 ; EFSA (European Food Safety Authority).…”

Section: Introductionmentioning

confidence: 99%

Lumpy skin disease in Kazakhstan

Orynbayev

Nissanova

Khairullin

et al. 2021

Trop Anim Health Prod

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This study describes the registration of the first cases of lumpy skin disease in July 2016 in the Republic of Kazakhstan. In the rural district of Makash, Kurmangazinsky district of Atyrau region, 459 cattle fell ill and 34 died (morbidity 12.9% and mortality 0.96%). To determine the cause of the disease, samples were taken from sick and dead animals, as well as from insects and ticks. LSDV DNA was detected by PCR in all samples from dead animals and ticks (Dermacentor marginatus and Hyalomma asiaticum), in 14.29% of samples from horseflies (Tabanus bromius), and in one of the samples from two Stomoxys calcitrans flies. The reproductive LSD virus was isolated from organs of dead cattle and insects in the culture of LT and MDBK cells. The virus accumulated in cell cultures of LT and MDBK at the level of the third passage with titers in the range of 5.5–5.75 log 10 TCID50/cm3. Sequencing of the GPCR gene allowed us to identify this virus as a lumpy skin disease virus.

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Section: Introductionmentioning

confidence: 99%

Lumpy skin disease in Kazakhstan

Orynbayev

Nissanova

Khairullin

et al. 2021

Trop Anim Health Prod

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“…The quick and uncomplicated setup can be easily integrated into existing monitoring protocols and may also provide diagnostic options for other important pathogens in zoological institutions. RPA has been adopted for other emerging and neglected infectious diseases [ 42 , 43 , 44 , 45 , 46 ]. Negative RPA results in our study were confirmed by negative results in real-time IS900 PCR.…”

Section: Discussionmentioning

confidence: 99%

Molecular and Serological Footprints of Mycobacterium avium Subspecies Infections in Zoo Animals

Roller

Hansen

Böhlken-Fascher

et al. 2020

Veterinary Sciences

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Background: Mycobacteria of the Mycobacterium avium complex (MAC) pose a significant risk to zoological collections. Mycobacterium avium subspecies paratuberculosis (MAP) is a member of MAC and the causative agent of Johne’s disease. Despite many reports in animals kept in zoological gardens, systemic surveillance has rarely been reported. Methods: In this study, archived serum samples collected from animal species at the Wilhelma Zoological and Botanical Gardens in Stuttgart, Germany, were screened for the presence of antibodies against MAC and MAP. In addition, molecular investigations were performed on necropsy, fecal, and environmental samples. Results: In total, 30/381 serum samples of various mammalian species were positive for MAC antibodies in ELISA, while one sample of a reticulated giraffe (Giraffa camelopardalis reticulata) was positive in MAP-specific ELISA. Samples from many species were positive in pan-Mycobacterium real-time PCR (40/43 fecal samples, 27/43 environmental samples, and 31/90 necropsy samples). Surprisingly, no sample was positive in the MAP-specific molecular assays. However, two environmental samples from primate enclosures were positive in Mycobacterium avium subspecies hominissuis (MAH)-specific real-time PCR. Conclusions: The results reveal serological indications of MAC infections in the zoological collection. However, the presence of a MAP-contaminated environment by a high-shedding individual animal or MAP-infected population is unlikely.

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“…However, the above methods have been either too time-consuming or require high-precision instruments to meet practical needs [ 19 , 20 ]. RPA isothermal amplification techniques can amplify nucleic acids and detect the products without a requirement of special instrument or complex operations [ 21 – 24 ]. It is worth mentioning that human body heat can indeed incubate RPA reactions under certain limit resources [ 25 ].…”

Section: Discussionmentioning

confidence: 99%

Development and evaluation of serotype-specific recombinase polymerase amplification combined with lateral flow dipstick assays for the diagnosis of foot-and-mouth disease virus serotype A, O and Asia1

et al. 2018

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BackgroundFoot-and-mouth disease (FMD) caused by foot-and-mouth disease virus (FMDV) is one of the most highly infectious diseases in livestock, and leads to huge economic losses. Early diagnosis and rapid differentiation of FMDV serotype is therefore integral to the prevention and control of FMD. In this study, a series of serotype-specific reverse transcription recombinase polymerase amplification assays combined with lateral flow dipstick (RPA-LFD) were establish to differentiate FMDV serotypes A, O or Asia 1, respectively.ResultsThe serotype-specific primers and probes of RPA-LFD were designed to target conserved regions of the FMDV VP1 gene sequence, and three primer and probe sets of serotype-specific RPA-LFD were selected for amplification of FMDV serotypes A, O or Asia 1, respectively. Following incubation at 38 °C for 20 min, the RPA amplification products could be visualized by LFD. Analytical sensitivity of the RPA assay was then determined with ten-fold serial dilutions of RNA of VP1 gene and the recombinant vector respectively containing VP1 gene from FMDV serotypes A, O or Asia1, the detection limits of these assays were 3 copies of plasmid DNA or 50 copies of viral RNA per reaction. Moreover, the specificity of the assay was assessed, and there was no cross reactions with other viruses leading to bovine vesicular lesions. Furthermore, 126 clinical samples were respectively detected with RPA-LFD and real-time PCR (rPCR), there was 98.41% concordance between the two assays, and two samples were positive by RPA-LFD but negative in rPCR, these were confirmed as FMDV-positive through viral isolation in BHK-21 cells. It showed that RPA-LFD assay was more sensitive than the rPCR method in this study.ConclusionThe development of serotype-specific RPA-LFD assay provides a rapid, sensitive, and specific method for differentiation of FMDV serotype A, O or Asia1, respectively. It is possible that the serotype-specific RPA-LFD assay may be used as a integral protocol for field detection of FMDV.

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Recombinase polymerase amplification assay for rapid detection of lumpy skin disease virus

Cited by 37 publications

References 33 publications

Lumpy skin disease in Kazakhstan

Lumpy skin disease in Kazakhstan

Molecular and Serological Footprints of Mycobacterium avium Subspecies Infections in Zoo Animals

Development and evaluation of serotype-specific recombinase polymerase amplification combined with lateral flow dipstick assays for the diagnosis of foot-and-mouth disease virus serotype A, O and Asia1

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