Evaluation of an incubation instrument-free reverse transcription recombinase polymerase amplification assay for rapid and point-of-need detection of canine distemper virus

Wang, Jianchang; Wang, Jinfeng; Li, Ruiwen; Shi, Ruihan; Liu, Libing; Yuan, Wanzhe

doi:10.1016/j.jviromet.2018.07.007

Cited by 18 publications

(13 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results proved that gene expression analysis test was an accurate, conclusive, confirmatory, qualitative test in diagnosis of CDV infection in live dogs as confirmed by many authors [Elia et al [18] and Shaw & Ihle [26]. Moreover that qRT-PCR was a sensitive anti mortem diagnostic technique especially reliable in sub acute and chronic stages of CDV infection [17,18,[28][29][30].…”

Section: So Ic Applied As a Rapid Field Test Reliable For Diagnosis supporting

confidence: 58%

Rapid Approaches for Diagnosis of Canine Distemper Virus in Live and Dead Dogs in Egypt

Awad

2019

NIDOC-ASRT

View full text Add to dashboard Cite

Paramyxovirridae, CDV is a negative single strand RNA. Enveloped, RNA of the virus encoding for 6 structural proteins: Fusion protein (F), Haemagglutinin protein (H), large polymerase (L), nucleoprotein capside (N), protein envelops matrix protein (M) and Phosphoprotein (P) [4,5]. Infection occurs via inhalation, CDV invade upper respiratory tract, lymphoid tissues (6), replicating inside macrophage and monocyte cells [7]. Incubation period varies from one to four weeks depend up on immune-status of dogs and strain virulence [6,8]. B ACkgRounD: Canine Distemper Virus infection (CDV) is a highly contagious disease of high morbidity and mortality rates in dogs. The causative virus is CDV which is a Morbillivirus, CDV is a pantropic virus characterized by multisystemic infection and high case fatality, with worldwide distribution, so rapid diagnosis and quarantine of the infected dogs with starting suitable treatment was required. The aim: this study aimed to achieve rapid diagnosis of canine distemper virus infection on ante mortem and post mortem aspects. Material and Methods: One healthy control disabled dog, 53 infected dogs with suggestive clinical signs for CDV infection were checked by (a) clinical examination; (b) Rapid immunochromatgrophy (IC) onconjunctival swabs (c) qRT-PCR on blood and tracheal exudates to confirm presence or absence of CDV, by expression analysis of CDV-F gene. Then all 53 examined dogs isolated and received supportive treatment but all died and disabled controlled one exposed to soft death. qRT-PCR were conducted on tissue samples from all 54 dogs for detection CDV-F gene expression values in different tissues.(d) Statistical analysis to study effect of sex and breed using Chi-square test, to evaluate sensitivity, specificity, accuracy, PPV, negative PV, respectively of used test and prevalence of CDV. Results: Clinical signs suggestive for CDV infection recorded in all 53 examined dogs, 24 of 54 dogs were positive for IC. Gene expression analysis test detected high values for CDV-F gene expression in tracheal exudates and blood samples of 36 live dogs, while the expression values were also high in tissue samples from different organs of 36 dead dogs. Statistical comparison of IC to qRT-PCR showed that values were 72%, 100%, 81.4%, 100% and 64.2 for sensitivity, specificity, accuracy, PPV, negative PV, respectively. No effect of sex , age, and breed on results using Chi-square test. Prevalence of CDV infection was 66% among population of this study. Conclusion: this study concluded that detection of clinical signs suggestive for CDV with application of IC and qRT-PCR together should be applied as rapid diagnosis on ante mortem level, while qRT-PCR could be used for rapid post mortem diagnosis of CDV infection.

show abstract

Section: So Ic Applied As a Rapid Field Test Reliable For Diagnosis supporting

confidence: 58%

Rapid Approaches for Diagnosis of Canine Distemper Virus in Live and Dead Dogs in Egypt

Awad

2019

NIDOC-ASRT

View full text Add to dashboard Cite

show abstract

“…Previously, Wang, et al developed a real-time RT-RPA assay for canine CDV, which can detect 31.8 copies per reaction of RNA transcripts in 12 min [ 16 ], but this assay is only employed in advanced laboratories because it requires sophisticated real-time PCR instruments. Subsequently, Wang, et al adjusted their design and developed a RT-RPA assay combined with lateral flow strips (LFS RT-RPA), which still had a high sensitivity of 94 copies RNA visible to the naked eye on the lateral flow strip [ 17 ]. The primers and probe sequences of LFS RT-RPA designed by Wang et al are containing 13 mismatch bases compared with giant panda/SX/2014 isolated from giant panda both belong to Asia-1 genotype.…”

Section: Discussionmentioning

confidence: 99%

Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda

Huang

Meng

et al. 2021

BMC Vet Res

View full text Add to dashboard Cite

Background Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). Results The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 101 copies/μl RNA transcripts and 100.5 TCID50 ml− 1 viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. Conclusions The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.

show abstract

“…In a comparison of real-time PCR, multiplex PCR and RT-PCR [25][26][27], the products of positive reactions of these assays were achieved through a series of temperature changes using specific instruments. But the emergence of RT-LAMP and Recombinase Polymerase Amplification (RPA) [28,29], based on isothermal amplification technology, makes sample detection faster, more economical and more operator friendly, as a consequence of the color-based visual results, based on color which provide a qualitative reaction that can be detected by the naked eye. Unfortunately, RPA technology uses a very complex enzyme reaction system resulting in higher detection costs [30] than with RT-LAMP.…”

Section: Discussionmentioning

confidence: 99%

Detection of melon necrotic spot virus by one-step reverse transcription loop-mediated isothermal amplification assay

Qiao¹,

Dai²,

Liu³

et al. 2020

PLoS ONE

View full text Add to dashboard Cite

Melon necrotic spot virus (MNSV) can cause significant economic losses due to decreased quality in cucurbit crops. The current study is the first to use reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of MNSV. A set of four LAMP primers was designed based on the coat protein gene sequence of MNSV, and a RT-LAMP reaction was successfully performed for 1 h at 62˚C. The results of RT-LAMP showed high specificity for MNSV and no cross-reaction with other viruses. Compared to traditional reverse transcription-PCR (RT-PCR), the RT-LAMP assay was 10 3-fold more sensitive in detecting MNSV. Due to its sensitivity, speed and visual assessment, RT-LAMP is appropriate for detecting MNSV in the laboratory.

show abstract

Evaluation of an incubation instrument-free reverse transcription recombinase polymerase amplification assay for rapid and point-of-need detection of canine distemper virus

Cited by 18 publications

References 24 publications

Rapid Approaches for Diagnosis of Canine Distemper Virus in Live and Dead Dogs in Egypt

Rapid Approaches for Diagnosis of Canine Distemper Virus in Live and Dead Dogs in Egypt

Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda

Detection of melon necrotic spot virus by one-step reverse transcription loop-mediated isothermal amplification assay

Contact Info

Product

Resources

About