Optimisation of reverse transcriptase loop-mediated isothermal amplification assay for rapid detection of Macrobrachium rosenbergii noda virus and extra small virus in Macrobrachium rosenbergii

Haridas, Divya V.; Pillai, Devika; Manojkumar, B.; Nair, C. Mohanakumaran; Sherief, P M

doi:10.1016/j.jviromet.2010.03.011

Cited by 21 publications

(18 citation statements)

References 23 publications

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“…LAMP is being used increasingly for rapid detection and typing of emerging viruses (10,12,20,21). The use of reverse transcription (RT)-LAMP with HNB dye for visual detection of pandemic influenza A H1N1 virus 2009 was developed recently in our laboratory (16).…”

Section: Discussionmentioning

confidence: 99%

Visual Detection of High-Risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye

et al. 2011

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A simple, rapid, sensitive, qualitative, colorimetric loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB) was established to detect high-risk human papillomavirus (HPV) genotypes 16, 18, 45, 52, and 58. All initial validation studies with the control DNA proved to be type specific. The colorimetric type-specific LAMP assay could achieve a sensitivity of 10 to 100 copies at 63°C for 65 min, comparable to that of real-time PCR. In order to evaluate the reliability of HPV type-specific LAMP, the assay was further evaluated with HPV DNAs from a panel of 294 clinical specimens whose HPV status was previously determined with a novel one-step typing method with multiplex PCR. The tested panel comprised 108 HPV DNA-negative samples and 186 HPV-DNA-positive samples of 14 genotypes. The results showed that the sensitivity of HPV type-specific LAMP for HPV types 16, 18, 45, 52, and 58 was 100%, 100%, 100%, 100%, and 100%, respectively, and the specificity was 100%, 98.5%, 100%, 98.8%, and 99.2%, respectively, compared with a novel one-step typing method with multiplex PCR. No cross-reactivity with other HPV genotypes was observed. In conclusion, this qualitative and colorimetric LAMP assay has potential usefulness for the rapid screening of HPV genotype 16, 18, 45, 52, and 58 infections, especially in resource-limited hospitals or rural clinics of provincial and municipal regions in China.It is well established that virtually all cervical cancers are caused by persistent high-risk oncogenic human papillomavirus (HPV) infections in the cervix. Thus, cervical screening could be improved by testing for the DNA of high-risk types of HPV as a primary screening tool (3, 9, 24). So far, at least 14 high-risk HPV genotypes have been shown to cause cervical cancer; those most important for cervical cancer are HPV types 16, 18, 45, 31, 33, 52, 58, and 35 (6, 7, 26). HPV16 is by far the predominant type, causing more than 50% of cancers; HPV18 follows, causing 14%; and HPV45 causes approximately 2.8%. These genotypes are highly prevalent in all regions of the world and account for approximately 70% of all cancers of the cervix (2, 13, 25). HPV types 52 and 58 are rare in Western countries; however, they are prevalent in Asian populations, especially in China (5,14,15,28,29).Technologies available for HPV genotyping vary by method and platform and may offer type-specific characterization for a multitude of different high-and low-risk HPV types. For example, The Qiagen Hybrid Capture 2 HPV DNA (HC2) test and the Roche Amplicor HPV test (Amplicor) detect the DNA of 13 high-risk HPV types. The Linear Array HPV genotyping test allows for the discrimination of 37 HPV genotypes. The PapilloCheck HPV screening test (Greiner Bio-One GmbH, Frickenhausen, Germany) is a PCR-based DNA microarray system for the detection and identification of 24 HPV genotypes (4, 7, 13, 25). However, these methods might not be suitable in primary clinical settings in developing countries or for field use, because of the sophisticated ...

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Section: Discussionmentioning

confidence: 99%

Visual Detection of High-Risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye

et al. 2011

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“…Haridas et al [3] and Pillai et al [12] have applied loopmediated isothermal amplification (LAMP) for rapid diagnosis of MrNV and XSV in the freshwater prawn. A set of four primers, two outer and two inner, have been designed separately for detection of MrNV and XSV.…”

Section: Diagnostic Methodsmentioning

confidence: 99%

White Tail Disease of Freshwater Prawn, Macrobrachium rosenbergii

Hameed¹,

Bonami

2012

Indian J. Virol.

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Macrobrachium rosenbergii is the most important cultured freshwater prawn in the world and it is now farmed on a large scale in many countries. Generally, freshwater prawn is considered to be tolerant to diseases but a disease of viral origin is responsible for severe mortalities in larval, post-larval and juvenile stages of prawn. This viral infection namely white tail disease (WTD) was reported in the island of Guadeloupe in 1995 and later in Martinique (FrenchWest Indies) in Taiwan, the People's Republic of China, India, Thailand, Australia and Malaysia. Two viruses, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus-like particle (XSV) have been identified as causative agents of WTD. MrNV is a small icosahedral non-enveloped particle, 26-27 nm in diameter, identified in the cytoplasm of connective cells. XSV is also an icosahedral virus and 15 nm in diameter. Clinical signs observed in the infected animals include lethargy, opaqueness of the abdominal muscle, degeneration of the telson and uropods, and up to 100 % within 4 days. The available diagnostic methods to detect WTD include RT-PCR, dot-blot hybridization, in situ hybridization and ELISA. In experimental infection, these viruses caused 100 % mortality in post-larvae but failed to cause mortality in adult prawns. The reported hosts for these viruses include marine shrimp, Artemia and aquatic insects. Experiments were carried out to determine the possibility of vertical transmission of MrNV and XSV in M. rosenbergii. The results indicate that WTD may be transferred from infected brooders to their offspring during spawning. Replication of MrNV and XSV was investigated in apparently healthy C6/36 Aedes albopictus and SSN-1 cell lines. The results revealed that C6/36 and SSN-1cells were susceptible to these viruses. No work has been carried out on control and prevention of WTD and dsRNA against protein B2 produced RNAi that was able to functionally prevent and reduce mortality in WTD-infected redclaw crayfish.

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“…However, the turbidity of the positive samples is not stable for a longer time and it should be judged soon after taking out of the samples from the water bath or from the thermal cycler. As a solution, adding SYBR Green and Ethidium Bromide to the tubes will provide enough time to monitor changes in the color of the tubes and to detect the positive samples under UV illumination as stressed by other workers [25,33]. However, using of toxic staining materials and UV irradiation would not be compatible with the main feature of this technique which is "safety".…”

Section: It Is a Rapid And A Highly Specific Methodsmentioning

confidence: 99%

“…Typically, in the recent decades the molecular techniques have been applied for discrimination of viruses and other plant pathogens. In this regards, several molecular methods have been developed for detection of the PLRV, including nucleic acid sequence-based amplification (NASBA) [16], Northern blotting [7], immunecapture RT-PCR (IC-RT-PCR) [3], reverse transcription polymerase chain reaction (RT-PCR) [16][17][18][19][20][21][22][23][24][25][26][27], Multiplex AmpliDet RNA [18], and real-time RT-PCR [1,19]. Although those methods are sensitive and specific, they are either complicated or expensive.…”

Section: Introductionmentioning

confidence: 99%

“…Up to now, numerous studies in both plants and animals have been reported to recognize pathogens by LAMP technique. For instance, RT-LAMP method were utilized for detection of Plasmodium falciparum gametocytes [9], Fusarium graminearum [23], Peste des petits ruminants virus (PPRV) [24], Potato virus Y (PVY) [10], Japanese yam mosaic potyvirus (JYMV) [22], Rabies virus (RABV) [19], Macrobrachium rosenbergii noda virus (MrNV) [25], Cymbidium *Corresponding author: Mohammad Amin Almasi, Department of Plant Biotechnology, Faculty of Agriculture, University of Zanjan, Zanjan, Iran, E-mail: aminalmasi66@gmail.com mosaic virus (CymMV) [26] and Potato Leafroll Virus (PLRV) [2,3]. As described earlier, some different visualizing systems are employed for verification of LAMP products.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Almasi¹

2012

J Plant Pathol Microb

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Loop-mediated isothermal amplification assay amplifies DNA/RNA with high specificity and sensitivity. In this study, we describe an optimized reverse transcription-LAMP assay for detection of Potato Leafroll Virus. Firstly, DAS-ELISA assay was performed to detect of the virus in a collection containing 40 suspicious samples. Lastly, two samples were detected as the positive samples. Then, the positive samples were verified by RT-PCR and RT-LAMP methods. Furthermore, the results demonstrated that the RT-LAMP assay was 40 times sensitive and 4 time faster compared to RT-PCR. RT-LAMP assay was accomplished in the water bath either frees from any thermal cycler machine or sophisticated laboratories facility. Moreover, in RT-LAMP reaction the positive samples were detected through turbidity which produced by magnesium pyrophosphate. Interestingly, the application of CaCl 2 instead of MgSO 4 which create calcium pyrophosphate in reaction could significantly increase both stability and concentration of turbidity. Consequently, it could be an interesting alternative to MgSO4. Overall, the newly developed RT-LAMP assay can be a sensitive, specific and low-cost method for early detection of Potato Leafroll Virus and also other viral plant pathogens.

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Optimisation of reverse transcriptase loop-mediated isothermal amplification assay for rapid detection of Macrobrachium rosenbergii noda virus and extra small virus in Macrobrachium rosenbergii

Cited by 21 publications

References 23 publications

Visual Detection of High-Risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye

Visual Detection of High-Risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye

White Tail Disease of Freshwater Prawn, Macrobrachium rosenbergii

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

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