Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Li, Zhili; Cai, Yuejia; Liang, Guozhi; El‐Ashram, Saeed; Mei, Minmin; Huang, Wenjing; Li, Xiaowen; Li, Wenfeng; He, Cheng; Huang, Shujian

doi:10.1038/s41598-018-32473-4

Cited by 18 publications

(21 citation statements)

References 34 publications

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“…Although the previous results suggest that HMGB1 has no effect on PRRSV and NDV replication, HMGB1 promotes virus-induced NF-κB activation and subsequent expression of inflammatory cytokines, enhances the efficiency of virus-induced inflammatory responses [16,31]. Thus, we tried to test the potential antiviral effect of duHMGB1 by comparing the viral load between duHMGB1 overexpression and knockdown in DEF cells, which showed an antiviral effect of duHMGB1 on DTMUV, NDRV and DPV (the infectious diseases caused by these three viruses has resulted in massive economic loss to the duck industry) [37][38][39]. Since the effect of duHMGB1 overexpression has been the most effective to inhibit viral replication of NDRV at all time-points (from 12 to 48 hpi), the immune response in DEF cells was subsequently studied after NDRV infection as an example.…”

Section: Discussionmentioning

confidence: 99%

High-mobility group box 1 protein (HMGB1) from Cherry Valley duck mediates signaling pathways and antiviral activity

Hou

Liu

Zhang

et al. 2020

Vet Res

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High-mobility group box 1 protein (HMGB1) shows endogenous damage-associated molecular patterns (DAMPs) and is also an early warning protein that activates the body's innate immune system. Here, the full-length coding sequence of HMGB1 was cloned from the spleen of Cherry Valley duck and analyzed. We find that duck HMGB1(duHMGB1) is mostly located in the nucleus of duck embryo fibroblast (DEF) cells under normal conditions but released into the cytoplasm after lipopolysaccharide (LPS) stimulation. Knocking-down or overexpressing duH-MGB1 had no effect on the baseline apoptosis rate of DEF cells. However, overexpression increased weakly apoptosis after LPS activation. In addition, overexpression strongly activated the IFN-I/IRF7 signaling pathway in DEF cells and significantly increased the transcriptional level of numerous pattern recognition receptors (PRRs), pro-inflammatory cytokines (IL-6, TNF-α), IFNs and antiviral molecules (OAS, PKR, Mx) starting from 48 h post-transfection. Overexpression of duHMGB1 strongly impacted duck virus replication, either by inhibiting it from the first stage of infection for novel duck reovirus (NDRV) and at late stage for duck Tembusu virus (DTMUV) or duck plague virus (DPV), or promoting replication at early stage for DTMUV and DPV infection. Importantly, data from duHMGB1 overexpression and knockdown experiments, time-dependent DEF cells transcriptional immune responses suggest that duHMGB1 and RIG-I receptor might cooperate to promote the expression of antiviral proteins after NDRV infection, as a potential mechanism of duHMGB1-mediated antiviral activity.

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Section: Discussionmentioning

confidence: 99%

High-mobility group box 1 protein (HMGB1) from Cherry Valley duck mediates signaling pathways and antiviral activity

Hou

Liu

Zhang

et al. 2020

Vet Res

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“…So, it is essential to develop molecular diagnosis techniques for the purpose of correct and rapid detection of viral pathogens in order to prevent further disease transmission or outbreaks. [11,13,14]. Although test results obtained through RT-PCR are reliable, this technique is too costly and time-consuming for companies or persons who have limited laboratory equipment, including a thermal cycler, and can only conduct mostly eld surveys [26].…”

Section: Discussionmentioning

confidence: 99%

“…At 37 ℃, the reaction can be completed within 20 minutes with only a pair of primers and a simple device such as water bath and heating block. Rapid and speci c detection of NDRV using the RT-LAMP procedure has also been reported for ducklings [14,15]. But RT-LAMP method requires high temperature (60-65 ℃), six primers and more di cult downstream analysis, such as cloning and direct sequencing, limitations that are overcome by the RT-RPA procedure.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the enzyme-linked immunosorbent assay (ELISA) have been reported to be effective in detecting NDRV [13], but it relies on high antibodies quality and speci city and easy to produce false positives. The reverse transcription loop-mediated isothermal ampli cation (RT-LAMP) was established to detect NDRV [14,15], which needs six primers, high temperature reaction (60-65 ℃) and long reaction time (50-65 min) makes it di cult as an on-site facilities rapid detection method [16].…”

Section: Introductionmentioning

confidence: 99%

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Rapid and Visual Detection of the Emerging Novel Duck Reovirus by Using a Specific and Sensitive Reverse Transcription Recombinase Polymerase Amplification Method

Wang

Zhang

Huang

et al. 2020

Preprint

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Background Duck spleen necrosis disease (DSND) caused by Novel Duck Reovirus (NDRV), is an emerging infectious disease that causes severely threaten to duck industry. Currently, the popular conventional PCR technique for detecting NDRV is time consuming. So, it is essential to develop a rapid and accurate molecular diagnosis techniques of viral pathogens for the purpose to prevent further disease transmission or outbreaks. Recombinase polymerase Amplification (RPA) is a new generation of simple, rapid and cost-effective molecular diagnosis technology, which has been applied to the molecular detection of various pathogens. Methods In our study, a simple, rapid and reliable detection method was developed target NDRV by an isothermal reverse transcription-recombinase polymerase amplification (RT-RPA). The RT-RPA primers were designed based on the S3 gene of NDRV, and a series of other waterfowl-origin pathogens were detected by RT-RPA. A total of 20 field and experimental infected samples were tested by RT-RPA and compared with the results of conventional RT-PCR and quantitative RT-PCR simultaneously. Results The RT-RPA method proved to be repeatable and could detect as little as 3.48 × 10− 6 ng/µl of the standard plasmid DNA inserted with the viral S3 gene. This was a 10 × higher sensitivity rate than that of conventional RT-PCR. The major advantage of this RT-RPA method is that it could be performed as an isothermal reaction at 37 ℃ and completed within 20 min. In addition, no cross-reactivity was detected with other waterfowl-origin viruses. Also, the amplified products could be visualized faster, without the gel electrophoresis, by adding the SYBR Green I and observing them under an ultraviolet light. Conclusions This newly developed RT-RPA method offers a simple, rapid and accurate for rapid detection of NDRV, which especially useful in on-site facilities and resource-limited areas.

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“…Conventional detection of MNSV is performed by enzyme-linked immunosorbent assay (ELISA) or reverse transcription-polymerase chain reaction (RT-PCR) [18], but these methods have the disadvantages of taking a long time to complete, and of requiring the availability of specialized equipment and expensive consumables. Recent research has used the reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to detect plant [19,20], animal [21,22] and human viruses [23,24]. This method has been shown to exhibit high sensitivity, reliability and rapid completion time; furthermore, it requires cheaper consumables and simpler equipment than the conventional RT-PCR detection method.…”

Section: Introductionmentioning

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

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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.

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Detection of Novel duck reovirus (NDRV) using visual reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Cited by 18 publications

References 34 publications

High-mobility group box 1 protein (HMGB1) from Cherry Valley duck mediates signaling pathways and antiviral activity

High-mobility group box 1 protein (HMGB1) from Cherry Valley duck mediates signaling pathways and antiviral activity

Rapid and Visual Detection of the Emerging Novel Duck Reovirus by Using a Specific and Sensitive Reverse Transcription Recombinase Polymerase Amplification Method

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

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