Evaluation of Recombinase Polymerase Amplification Assay for Detecting<i>Meloidogyne javanica</i>

Chi, Yuan-Kai; Zhao, Wei; Ye, Meng-di; Ali, Farman; Wang, Tao; Ran, Qi

doi:10.1094/pdis-07-19-1473-re

Cited by 27 publications

(14 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For desinging the species-specific M. javanica RPA primers and probe we used a genome fragment firstly proposed by Randig et al (2002) and then applied for RPA assays by Ju et al (2019) and Chi et al (2020) . In our RPA assay the forward species-specific RPA primer proposed by Ju et al (2019) and a new reverse primer were used.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, RPA assays with a gel visualization of amplicons for species diagnostics of M. javanica, M. arenaria, M. incognita and M. enterolobii have also been developed by Ju et al (2019) . Chi et al (2020) designed a rapid RPA assay for visual detection of M. javanica using SYBR Green and lateral flow dipsticks (LF-RPA) at endpoint. RPA assays for detection of the northern RKN, M. hapla were also developed by Song et al (2021) and Subbotin and Burbridge (2021) .…”

mentioning

confidence: 99%

See 1 more Smart Citation

Recombinase Polymerase Amplification assays for detection of the major tropical root-knot nematodes

Subbotin

Burbridge

2021

Journal of Nematology

View full text Add to dashboard Cite

Detection of root-knot nematodes (RKN) in soil and plant samples is crucial to prevent its spread and select effective control measures. In this study, Recombinase Polymerase Amplification (RPA) assays using lateral flow dipsticks (LF-RPA) and real-time fluorescence detection (real-time RPA) were developed to detect the RKN species from tropical complex using a group-specific primer-probe set and Meloidogyne javanica using a species-specific primer-probe set. The results of the real time RPA assays in series of crude nematode extracts showed reliable detection within 16 min with a sensitivity of 1/100 of a second-stage juvenile in a reaction tube. The results of the LF-RPA assays showed reliable detection within 30 min with a sensitivity of 1/20 to 1/100 of a second-stage juvenile and 1/10 of a female in a reaction tube. Real-time RPA and LF-RPA assays are highly specific and can identify their target DNA in mixtures with other nematodes and plant tissues. LF-RPA assay has great potential for diagnosing RKN in the lab, field or in areas with a minimal laboratory infrastructure.

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Recombinase Polymerase Amplification assays for detection of the major tropical root-knot nematodes

Subbotin

Burbridge

2021

Journal of Nematology

View full text Add to dashboard Cite

show abstract

“…Different RPA-based assays have been developed to successfully detect different pathogens from different specimen [18][19][20][21]. In the wake of the COVID-19 pandemic, RT-RPA has been adapted and used to amplify the envelope protein (E) and RNA-dependent RNA polymerase (RdRP) genes of SARS-CoV-2 successfully, showing a good sensitivity and specificity compared with RT-PCR [22].…”

Section: Recombinase Polymerase Amplificationmentioning

confidence: 99%

Rapid, Cheap, and Effective COVID-19 Diagnostics for Africa

et al. 2021

View full text Add to dashboard Cite

Background: Although comprehensive public health measures such as mass quarantine have been taken internationally, this has generally been ineffective, leading to a high infection and mortality rate. Despite the fact that the COVID-19 pandemic has been downgraded to epidemic status in many countries, the real number of infections is unknown, particularly in low-income countries. However, precision shielding is used in COVID-19 management, and requires estimates of mass infection in key groups. As a result, rapid tests for the virus could be a useful screening tool for asymptomatic virus shedders who are about to come into contact with sensitive groups. In Africa and other low- and middle-income countries there is high rate of COVID-19 under-diagnosis, due to the high cost of molecular assays. Exploring alternate assays to the reverse transcriptase polymerase chain reaction (RT-PCR) for COVID-19 diagnosis is highly warranted. Aim: This review explored the feasibility of using alternate molecular, rapid antigen, and serological diagnostic assays to accurately and precisely diagnose COVID-19 in African populations, and to mitigate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RT-PCR diagnostic challenges in Africa. Method: We reviewed publications from internet sources and searched for appropriate documents available in English. This included Medline, Google Scholar, and Ajol. We included primary literature and some review articles that presented knowledge on the current trends on SARS-CoV-2 diagnostics in Africa and globally. Results: Based on our analysis, we highlight the utility of four different alternatives to RT-PCR. These include two isothermal nucleic acid amplification assays (loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA)), rapid antigen testing, and antibody testing for tackling difficulties posed by SARS-CoV-2 RT-PCR testing in Africa. Conclusion: The economic burden associated COVID-19 mass testing by RT-PCR will be difficult for low-income nations to meet. We provide evidence for the utility and deployment of these alternate testing methods in Africa and other LMICs.

show abstract

“…Using a lateral flow dipstick (LFD) strip, RPA amplicons can be observed directly with the naked eye, making it suitable for application in the field [13]. To date, RPA has been successfully used to detect a variety of plant parasitic nematodes [16], including Meloidogyne spp., such as M. incognita, M. arenaria, M. javanica, M. enterolobii [17], M. javanica [18], and M. enterolobii [19]. In addition, Cha et al reported the application of RPA for detecting Bursaphelenchus xylophilus [20,21].…”

Section: Introductionmentioning

confidence: 99%

Rapid and Visual Detection of Heterodera schachtii Using Recombinase Polymerase Amplification Combined with Cas12a-Mediated Technology

Yao

Peng

Jiang

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

Heterodera schachtii is a well-known cyst nematode that causes serious economic losses in sugar beet production every year. Rapid and visual detection of H. schachtii is essential for more effective prevention and control. In this study, a species-specific recombinase polymerase amplification (RPA) primer was designed from a specific H. schachtii sequence-characterized amplified region (SCAR) marker. A band was obtained in reactions with DNA from H. schachtii, but absent from nontarget cyst nematodes. The RPA results could be observed by the naked eye, using a lateral flow dipstick (LFD). Moreover, we combined CRISPR technology with RPA to identify positive samples by fluorescence detection. Sensitivity analysis indicated that 10−4 single cysts and single females, 4−3 single second-stage juveniles, and a 0.001 ng genomic DNA template could be detected. The sensitivity of the RPA method for H. schachtii detection is not only higher than that of PCR and qPCR, but can also provide results in <1 h. Consequently, the RPA assay is a practical and useful diagnostic tool for early diagnosis of plant tissues infested by H. schachtii. Sugar beet nematodes were successfully detected in seven of 15 field sugar beet root samples using the RPA assay. These results were consistent with those achieved by conventional PCR, indicating 100% accuracy of the RPA assay in field samples. The RPA assay developed in the present study has the potential for use in the direct detection of H. schachtii infestation in the field.

show abstract

Evaluation of Recombinase Polymerase Amplification Assay for DetectingMeloidogyne javanica

Cited by 27 publications

References 31 publications

Recombinase Polymerase Amplification assays for detection of the major tropical root-knot nematodes

Recombinase Polymerase Amplification assays for detection of the major tropical root-knot nematodes

Rapid, Cheap, and Effective COVID-19 Diagnostics for Africa

Rapid and Visual Detection of Heterodera schachtii Using Recombinase Polymerase Amplification Combined with Cas12a-Mediated Technology

Contact Info

Product

Resources

About