Application of loop-mediated isothermal amplification assay in the detection of herpesvirus of turkey (FC 126 strain) from chicken samples in Nigeria

Adedeji, Adeyinka Jeremy; Abdu, Paul Ayuba; Luka, Pam Dachung; Owoade, A. A.; Joannis, Tony M.

doi:10.14202/vetworld.2017.1383-1388

Cited by 5 publications

(9 citation statements)

References 30 publications

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“…The result turnover time for cLAMP was within 30 min revealing that the method is faster than the conventional RT-PCR which requires approximately 4-5 h for endpoint gel electrophoresis. This agrees with the report of other authors [ 19 , 23 , 24 ] indicating the robustness of the LAMP technique. It has been reported that LAMP amplifies the target viral genome within 1 h [ 22 ].…”

Section: Discussionsupporting

confidence: 93%

“…The finding in this study agrees with the reports of previous authors [ 19 , 21 , 24 ] who employed the RT-LAMP technique to detect PPR viral genome in China, India, and Pakistan respectively. Furthermore, the LAMP assay was successfully used to detect herpesvirus of turkey from chicken samples in Nigeria [ 23 ]. The cLAMP kit also detected PPR viral genome in apparently healthy sheep and goats, indicating that the kit can be used to detect animals incubating the virus but without clinical signs.…”

Section: Discussionmentioning

confidence: 99%

“…[ 24 ] who detected PPR viral genome using RT-LAMP. However, RT-LAMP assay reported by the previous authors [ 19 , 21 , 23 ] used purified RNA/DNA. Isolation of purified RNA/DNA essentially demands a sterile environment and sophisticated laboratory equipment and conditions (like an expensive and lengthy process of RNA extraction), and this limits the application of RT-PCR and RT-LAMP in the field and poorly equipped laboratories in resource-poor country.…”

Section: Discussionmentioning

confidence: 99%

“…The troubles/problems of optimization of the RT-LAMP reagent are not a feature in cLAMP [ 29 ], as the cLAMP mix is an optimized formulation of Bst 2.0 WarmStart DNA polymerase and WarmStart RTx in a special low buffer reaction. The previous reports on RT-LAMP required an addition of a fluorescent dye like SYBR green or Eva green into the LAMP reaction [ 17 , 19 , 24 ] for end-product evaluation with the naked eyes or the use of 2% agarose gel electrophoresis stained with ethidium bromide [ 19 , 23 , 24 ] where ladder-like bands confirm positive samples. These expensive dyes consequently increase the cost of the RT-LAMP assay, while the electrophoresis procedure and gel imaging lengthen the duration of the RT-LAMP procedures.…”

Section: Discussionmentioning

confidence: 99%

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Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

et al. 2020

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Background and Aim: Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study aimed to compare the application of a commercial colorimetric loop-mediated isothermal amplification (cLAMP) kit and reverse transcriptase-polymerase chain reaction (RT-PCR) in the diagnosis of PPR in sheep and goats in Southeast Nigeria. Materials and Methods: Nasal swab samples were collected from West African Dwarf sheep and goats showing clinical signs suggestive of PPR (n=80) and those without any clinical signs (n=140) of the disease. The diagnosis was achieved through detection of PPR viral genome in the samples using a cLAMP kit and RT-PCR. cLAMP assay was done directly on nasal swab samples without ribosomal nucleic acid extraction. A set of six primers targeting the matrix gene protein was used for the cLAMP assay. Results: PPR viral genome was detected by both cLAMP and RT-PCR in 51 (63.8%) of the 80 samples from sheep and goats with signs suggestive of PPR while 14 (10%) of those without signs tested positive for PPR by both assay methods. There was a 100% agreement in the cLAMP and RT-PCR results. However, cLAMP was a faster, easier, and less expensive method compared to RT-PCR. Conclusion: The cLAMP assay demonstrates the potential for a point of care diagnosis in the field and a valuable diagnostic tool in areas with poor electricity supply as well as in a less equipped diagnostic laboratory. Since the reagents are affordable, cLAMP can be a diagnostic tool of choice in the detection and surveillance of PPR virus in countries with limited resources.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

et al. 2020

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“…Currently, LAMP has been widely used for infectious disease diagnostic applications where the researcher's main objective is to correctly identify positive and negative LAMP reactions to identify a specific disease . A positive LAMP reaction indicates that the target DNA sequence, representing a disease or other diagnosis such as human influenza viruses H1 or H3 , has been amplified, and therefore identified in that sample.…”

Section: Introductionmentioning

confidence: 99%

Mathematical model to reduce loop mediated isothermal amplification (LAMP) false‐positive diagnosis

2019

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Loop mediated isothermal amplification (LAMP) is a nucleic acid amplification technique performed under isothermal conditions. The output of this amplification technique includes multiple different sizes of deoxyribonucleic acid (DNA) structures which are identified by a banding pattern on gel electrophoresis plots. Although this is a specific amplification technique, the complexity of the primer design and amplification still lead to the issue of obtaining false‐positive results, especially when a positive reading is determined solely by whether there is any banding pattern in the gel electrophoresis plot. Here, we first performed extensive LAMP experiments and evaluated the DNA structures using microchip electrophoresis. We then developed a mathematical model derived from the various components that make up an entire LAMP structure to predict the full LAMP structure size in base pairs. This model can be implemented by users to make predictions for specific, DNA size dependent, banding patterns on their gel electrophoresis plots. Each prediction is specific to the target sequence and primers used and therefore reduces incorrect diagnosis errors through identifying true‐positive and false‐positive results. This model was accurately tested with multiple primer sets in house and was also translatable to different DNA and RNA types in previously published literature. The mathematical model can ultimately be used to reduce false‐positive LAMP diagnosis errors for applications ranging from tuberculosis diagnostics to E. coli to numerous other infectious diseases.

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Molecular characterization and phylogenetic analysis of Marek’s disease virus in chickens from Ogun State, Nigeria

Oluwayinka,

Otesile,

Oni

et al. 2023

Avian Pathology

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Application of loop-mediated isothermal amplification assay in the detection of herpesvirus of turkey (FC 126 strain) from chicken samples in Nigeria

Cited by 5 publications

References 30 publications

Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

Mathematical model to reduce loop mediated isothermal amplification (LAMP) false‐positive diagnosis

Molecular characterization and phylogenetic analysis of Marek’s disease virus in chickens from Ogun State, Nigeria

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