Detection of Babesia gibsoni in dogs by combining recombinase polymerase amplification (RPA) with lateral flow (LF) dipstick

Cui, Jie; Zhao, Yangnan; Sun, Yali; Yu, Liang; Liu, Qin; Zhan, Xueyan; Li, Muxiao; He, Li; Zhao, Junlong

doi:10.1007/s00436-018-6104-3

Cited by 12 publications

(9 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another end-point detection format, real-time recombinase polymerase amplification (RT-RPA) assay, employs a device to collect a fluorescence signal. Several RPA detection methods have been developed so far; in addition to animal virus, plant virus, fungi, bacteria, mycoplasma, and parasite RPA detection methods have been developed in recent years (Cui and Zhao, 2018;Karakkat and Hockemeyer, 2018;Zeng and Luo, 2019;Zhao and Hou, 2018;Hu and Zhong, 2019). Although the RPA technology has not been promoted on the market, some reports have verified its capabilities and its potential to eclipse PCR for further revolutions the life sciences (Li and Macdonald, 2019).…”

Section: Discussionmentioning

confidence: 99%

Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

Wang

Hou

2019

Journal of Virological Methods

View full text Add to dashboard Cite

Feline panleukopenia caused by feline parvovirus (FPV), a single-stranded DNA virus, is typically highly contagious and often presents with lethal syndrome. The broad spectrum of possible hosts suggests its potential for transmission from animal to person through close contact with pets. FPV thus serves as an example of the importance of new rapid point-of-care field diagnostic tools for the control and prevention of transmission, especially among rare wild animals and pet cats. Recombinase polymerase amplification (RPA), as a real-time and isothermal method, could be a more affordable alternative to PCR when combined with a lateral flow dipstick (LFD) indicator. In this study, we report a novel FPV lateral flow dipstick RPA (LFD-RPA) instant detection method capable of detecting a range of different FPV strains. The LFD-RPA assay consists of specific primers, probe, and nucleic acid strip. It is capable of detecting 10 2 copies of target nucleic acid per reaction, which is one order of magnitude higher than the sensitivity of traditional PCR. The most suitable reaction conditions for this assay are at 38 ℃ for 15 min. This paper develops an efficient visual detection system that can eliminate the need for professional staff and expensive and sophisticated equipment for field detection.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

Wang

Hou

2019

Journal of Virological Methods

View full text Add to dashboard Cite

show abstract

“…By using this sensitive QubeMDx PCR system, the detection limit of 0.002% of parasitemia (94 parasites/μL of blood) allows for early diagnosis of infected dogs. Although greater detection limit was previously demonstrated by the real-time PCR (9 parasites/μL of blood) [15], recombinase polymerase amplification-lateral flow (LF-RPA) dipstick method (0.5 parasites/μL of blood) [5], loop-mediated isothermal amplification (LAMP)(0.0000135% of parasitemia) [18], the differences can be attributed to the use of rapid DNA extraction procedures in the QubeMDx system, leading to a compromised DNA yield.…”

Section: Discussionmentioning

confidence: 99%

“…In addition to a low detection limit, a 100% of sensitivity and specificity for B. gibsoni detection was achieved by this QubeMDx PCR based on real-time PCR confirmed positive/negative samples, while previously 86.67 and 57.33% of positive rate were obtained by the LF-RPA dipstick method [5] and the LAMP method [18], respectively, evaluated from clinically suspected dogs. Furthermore, the internal control included in each reaction of QubeMDx PCR provides an additional source for quality control.…”

Section: Discussionmentioning

confidence: 99%

“…Babesia gibsoni ( B. gibsoni ) is a widespread tick-transmitted protozoan blood parasite that was first identified in 1910 in dogs and jackals from India and now prevalent in many regions, including Taiwan [1, 2] and other Asian countries such as Japan [3], Korea [4], China [5], Bangladesh [6], India [7] and Malaysia [8]. B. gibsoni infection is frequently found in companion dogs and presents a serious clinical problem in the world.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A novel PCR-based point-of-care method enables rapid, sensitive and reliable diagnosis of Babesia gibsoni infection in dogs

Liu

Chi²,

Chang³

et al. 2019

BMC Vet Res

View full text Add to dashboard Cite

BackgroundBabesia gibsoni (B. gibsoni) is an intraerythrocytic protozoan parasite of dogs that causes fever and hemolytic illness. A timely diagnosis is essential for the disease management.ResultsHere, we report a QubeMDx PCR system which enables a rapid, sensitive and reliable diagnosis of B. gibsoni near the dog patient. Within 30 min, this diagnostic assay was able to detect as low as 0.002% parasitemia of the dog blood. Using clinical samples, this new assay was validated to demonstrate 100% agreement with real-time PCR.ConclusionsThis novel diagnostic method provides a reliable point-of-care test to assist in the identification of B. gibsoni.

show abstract

“…12 This versatile PCR replacement finds its applications in different fields as diverse as biodefence, 13 agriculture, 14 water testing, 15 and food testing, 16 as well as medical diagnostics. 17 Moreover, this isothermal technique can be adapted to a range of microfluidic 18 and lateral flow systems 19 to achieve point-of-care (POC) testing with superb sensitivity. However, despite its fast assay speed and simple experimental procedure, RPA has been suggested as poorly suited for mutation detection.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Detection of Low-Abundance BRAF V600E Mutation in Fine-Needle Aspiration Samples by Zip Recombinase Polymerase Amplification

et al. 2021

View full text Add to dashboard Cite

Papillary thyroid carcinoma (PTC) is the most common thyroid cancer with high incidence in endocrine tumors, which emphasizes the significance of accurate diagnostics. Still, the commonly used cytological method (fine-needle aspiration (FNA) cytology) and molecular diagnostic methods (such as PCR and sequencing) are limited in terms of diagnostic time, sensitivity, and user-friendliness. In this study, we introduce a novel Zip recombinase polymerase amplification (Z-RPA) strategy to efficiently detect rare mutant alleles in PTC fine-needle aspiration samples, which is sensitive, fast, and simple to manipulate. Using Zip nucleic acid (ZNA) probes to clamp the mutation region, the phi 29 polymerase could selectively displace mismatched ZNA probes and start amplification, while leaving complementary ZNA probes untouched and blocking amplification according to genotype. We demonstrated the good sensitivity and specificity of this strategy with optimized conditions and design, which enabled detection of BRAF V600E mutation in a total 4 ng of genomic DNA within 40 min (≈1 copy). Robust behavior in clinical specimen analysis was also demonstrated. The Z-RPA strategy provides a pragmatic approach to rapidly, sensitively, and easily detect BRAF V600E mutation in clinical fine-needle aspiration samples, which is a promising method for early cancer diagnosis and treatment guideline.

show abstract

Detection of Babesia gibsoni in dogs by combining recombinase polymerase amplification (RPA) with lateral flow (LF) dipstick

Cited by 12 publications

References 34 publications

Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

A novel PCR-based point-of-care method enables rapid, sensitive and reliable diagnosis of Babesia gibsoni infection in dogs

Highly Sensitive Detection of Low-Abundance BRAF V600E Mutation in Fine-Needle Aspiration Samples by Zip Recombinase Polymerase Amplification

Contact Info

Product

Resources

About