TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically

Gong, Zhuandı; Shen, Xıaoyun; Liang, Huipeng; Geng, Jiawei; Wei, Shaozhong

doi:10.3906/vet-1907-80

Cited by 4 publications

(8 citation statements)

References 19 publications

(29 reference statements)

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“…Serological methods include enzyme-linked immunosorbent assays (ELISA), fluorescent antibody techniques (FA), virus neutralization tests, and immunocolloidal gold techniques [42,43]. On the other hand, molecular biology approaches comprise techniques such as PCR, qPCR, multiplex PCR, and loop-mediated isothermal amplification (LAMP) [44][45][46]. Despite their prevalent use, these methods have limitations.…”

Section: Discussionmentioning

confidence: 99%

Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus

Li,

Pan,

et al. 2024

Transboundary and Emerging Diseases

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Bovine coronavirus (BCoV) is a notable pathogen affecting newly born calves and adult cattle, increasing mortality rates among calves and reducing productivity in meat and dairy industries, thereby causing substantial economic losses. Current primary laboratory methods for detecting BCoV include RT-PCR assay, real-time RT-PCR assay, and ELISA. However, these methods are time-consuming, require specialized technicians, and necessitate a laboratory environment. Consequently, there is an urgent need for a rapid, sensitive, and easy to use diagnostic method to detect BCoV. This study introduces two innovative protocols: the real-time fluorescent reverse transcription recombinase-aided amplification (RT-RAA) and the test strip RT-RAA (RT-RAA-LFD). Our results indicate that real-time RT-RAA can complete the reaction in 20 min at 39°C, while RT-RAA-LFD can achieve detection in just 17.5 min at 35°C. These new approaches offer higher specificity, with no cross-reactivity to other viruses, and significantly enhanced sensitivity compared to existing methods (1.46 × 101 and 1.46 × 102 copies/μL, respectively). We evaluated the performance of our methods using 242 clinical samples, and compared with RT-PCR and RT-qPCR. Both real-time RT-RAA and RT-qPCR yielded similar detection rates, the detection rate of RT-RAA-LFD was better than RT-PCR. The RT-RAA methods developed in this study effectively overcome the limitations associated with both RT-PCR and RT-qPCR by offering advantages including a single, low reaction temperature that allows for room temperature operation. Both methods boast shorter reaction times, simpler and more portable instrumentation, as well as reduced technical and environmental demands. Generally, both RT-RAA methods established in this study offer new avenues for the rapid detection of BCoV, contributing significantly to the monitoring, prevention, and control of the disease in global bovine industry.

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Section: Discussionmentioning

confidence: 99%

Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus

Li,

Pan,

et al. 2024

Transboundary and Emerging Diseases

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“…The researchers made use of primers targeting the N and S genes of the viruses (the S gene was used for BCoV) within the same reaction and reported high sensitivity and specificity for these agents with their assay. Similarly, another triplex qPCR was developed by scientists in China for the detection of bovine parvovirus (BPV), BCoV, and bovine parainfluenza virus (BPIV), all of which are responsible for gastrointestinal and respiratory disease in cattle [56]. The triplex PCR was seen to yield a diagnostic sensitivity 1000 times greater than the conventional PCR and high sensitivity with a limit of detection of 2.0 × 10 2 RNA copies/μL and improved efficiency in clinical samples.…”

Section: Multiplex Pcrmentioning

confidence: 99%

Section: Multiplex Pcrmentioning

confidence: 99%

“…As an important pathogenic agent responsible for diarrhea and gastrointestinal disease in cattle, BCoV has been included in a number of studies that developed multiplex PCRs targeting enteric pathogens in calves and adult cattle [56][57][58][59][60][61][62]. Furthermore, BCoV is also an important agent in the BRDC and as such is also included in multiplex PCRs and screens targeting respiratory pathogens [43,56,63]. Therefore, there is great interest in developing effective assays that can screen and detect BCoV along with a number of other respiratory and gastrointestinal pathogens in order to save resources and assist producers and veterinarians.…”

Section: Multiplex Pcrmentioning

confidence: 99%

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Advances in Laboratory Diagnosis of Coronavirus Infections in Cattle

van den Hurk,

Regmi,

Naikare

et al. 2024

Preprint

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Coronaviruses cause infections in humans and diverse species of animals and birds with a global distribution. Bovine coronavirus (BCoV) produces predominantly two forms of disease in cattle, a respiratory form, and a gastrointestinal form. All age groups of cattle are affected by the respiratory form of coronavirus whereas the gastroenteric form causes neonatal diarrhea or calf scour in young cattle and winter dysentery in adult cattle. The tremendous impacts of bovine respiratory disease and the associated losses are well-documented and underscores the importance of this pathogen. Beyond this, studies have demonstrated significant impacts on milk production associated with outbreaks of winter dysentery, with up to a 30% decrease in milk yield. Research data so far have shown that the viral structure or strain is not a determinant of tissue tropism and clinical form of the disease. In North America, BCoV was identified for the first time in 1972, and it continues to be a significant economic concern for the cattle industry. The virus is classified under Coronaviridae family, Betacoronavirus genus and Betacorovirus 1 species. A number of conventional and molecular diagnostic assays are available for the detection of BCoV from clinical samples. Conventional assays for BCoV detection include virus isolation, which is challenging from clinical samples, electron microscopy, fluorescent antibody assays, and various immunoassays. Molecular tests are mainly based on nucleic acid detection and predominantly include conventional and real-time polymerase chain reaction (PCR) assays. Isothermal amplification assays and genome sequencing have also been used increasingly in recent years for the detection and identification of BCoV. Loop-mediated isothermal amplification, helicase-dependent amplification, recombinase polymerase amplification, and multienzyme isothermal rapid amplification are some examples of the isothermal amplification assays for used BCoV. Point-of-care testing has gained some popularity recently as they can offer user-friendly testing on the farm (barn-side). The present study reviewed literature on coronavirus infections in cattle from the last three and a half decades and presents information mainly on the current and advancing diagnostics in addition to epidemiology, clinical presentations, and the impact of the disease on the cattle industry.

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“…Traditionally, BPV detection is mainly based on serological, etiological, and molecular methods. Most traditional methods require a long time and special laboratory diagnosis equipment ( Wang et al, 2019a ; Gong et al, 2020 ). Therefore, a rapid, specific, and convenient method for field detection is of practical significance for the prevention and control of BPV infections.…”

Section: Introductionmentioning

confidence: 99%

Development of a colloidal gold immunochromatographic strip with enhanced signal for the detection of bovine parvovirus

Jin

Zhang

et al. 2023

Front. Microbiol.

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Bovine parvovirus (BPV) is a pathogen responsible for respiratory and digestive tract symptoms in calves and abortion and stillbirth in pregnant cows. In this study, we developed a colloidal gold immunochromatographic (GICG) strip with an enhanced signal for detecting BPV according to the double-antibody sandwich principle and an enzyme-based signal amplification system to amplify the signal. This system utilizes horseradish peroxidase reacting with a substrate solution containing 3,3′,5,5′-tetramethylbenzidine and dextran sulfate to obtain insoluble blue products on the test and control lines. We optimized different reaction conditions, including the amount of monoclonal antibodies (mAbs), pH of the colloidal gold solution, coating solution, blocking solution, sample pad treatment solution, antibody concentration in the control line, and antibody concentration in the detection line. The sensitivity of the signal-enhanced GICG strip showed that the minimum amount for detecting BPV was 102 TCID50, 10 times higher than that of the traditional GICG strip. The results of the specificity test showed that the signal-enhanced GICG strip had no cross-reactivity with BRV, BVDV, or BRSV. The results of the repeatability test showed that the coefficient of variation between and within batches was less than 5%, showing good repeatability. Moreover, for validation, PCR and the signal-enhanced GICG strip were used to detect 280 clinical bovine fecal samples. The concordance rate compared with PCR was 99.29%. Hence, the developed strip exhibited high sensitivity and specificity for the detection of BPV. Therefore, this strip could be a rapid, convenient, and effective method for the diagnosis of BPV infection in the field.

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TaqMan probe qRT-PCR detects bovine parvovirus and applies clinically

Cited by 4 publications

References 19 publications

Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus

Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus

Advances in Laboratory Diagnosis of Coronavirus Infections in Cattle

Development of a colloidal gold immunochromatographic strip with enhanced signal for the detection of bovine parvovirus

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