Direct detection of human herpesvirus 6 DNA in serum by the loop-mediated isothermal amplification method

Ihira, Masaru; Akimoto, Shiho; Miyake, Fumi; Fujita, Ayano; Sugata, Ken; Suga, S.; Ohashi, Masahiro; Nishimura, Naoko; Ozaki, Takao; Asano, Yoshizo; Yoshikawa, Tetsushi

doi:10.1016/j.jcv.2007.02.001

Cited by 53 publications

(43 citation statements)

References 34 publications

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“…The heat-treatment LAMP assay without DNA extraction could detect the DNA from a 10 -6 virus dilution, which was 100 and 10 times more sensitive than results obtained with the non heat-treatment LAMP assay without DNA extraction and the original LAMP assay with DNA extraction, respectively. As previously reported [4,15,16], the addition of a heat-treatment step increased the sensitivity of the LAMP assay. The detection limits of the LAMP assays without DNA extraction were the same as in saline, PBS and a transport medium containing a nasal swab (Table 1).…”

supporting

confidence: 69%

“…Kaneko et al [6] have reported that the sensitivity of the LAMP assay is less affected by the various components of the clinical samples than the PCR assay. It has also been reported that herpes simplex virus and human herpesvirus 6 DNA can be directly detected in clinical samples by the LAMP assay without DNA extraction [3][4][5].…”

mentioning

confidence: 99%

“…Several groups have reported that an additional heat-treatment step increases the sensitivity of the LAMP assay [4,15,16]. Therefore, to increase the sensitivity, we added a heat-treatment step to the LAMP assay in this study.…”

mentioning

confidence: 99%

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Direct Detection of Equine Herpesvirus Type 1 DNA in Nasal Swabs by Loop-Mediated Isothermal Amplification (LAMP)

Nemoto

Ohta

Tsujimura

et al. 2011

The Journal of Veterinary Medical Science

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ABSTRACT. We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.KEY WORDS: equine herpesvirus type 1 (EHV-1), heat treatment, loop-mediated isothermal amplification (LAMP).

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supporting

confidence: 69%

mentioning

confidence: 99%

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Direct Detection of Equine Herpesvirus Type 1 DNA in Nasal Swabs by Loop-Mediated Isothermal Amplification (LAMP)

Nemoto

Ohta

Tsujimura

et al. 2011

The Journal of Veterinary Medical Science

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“…34 The LAMP reaction using serial dilutions of plasmid showed a high sensitivity comparable to other LAMP assays. 28,29,31 However, the sensitivity of the assay using stool specimens will also be determined by the capacity of the extraction method to retain purified DNA, inactivate DNases, and remove polymerase inhibitors, such as bile salts and plant-based polysaccharides. 35,36 In S. stercoralis infection where low numbers of larvae may have an uneven distribution in the stool, the amount of stool used in the extraction method may also be significant.…”

Section: Discussionmentioning

confidence: 99%

“…Before the addition of 8 units (1 μL) of Bst (large fragment) DNA polymerase (New England Biolabs), the reaction solution and DNA extract was heated to 95 C for 5 minutes and cooled to room temperature, to denature the template DNA and facilitate primer binding to the target sequences. 28 After pulse centrifugation, DNA polymerase was added to the tubes. The LAMP reaction occurred over 60 minutes by heating the solution to 60 C, a temperature that produced the shortest reaction time.…”

Section: Methodsmentioning

confidence: 99%

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye

Watts

James

Sultana

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to 10 copies of target sequence in a plasmid and up to a 10 -2 dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation.

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Sequence‐specific detection method for reverse transcription, loop‐mediated isothermal amplification of HIV‐1

Curtis

Rudolph

Owen

2009

Journal of Medical Virology

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HIV diagnosis at the point-of-care or in resource-limited settings poses considerable challenges due to time and cost limitations. Currently, nucleic acid-based tests are the only reliable method for diagnosing recent infections during the window period post-infection and pre-seroconversion, but these tests are only suitable for well-equipped laboratory settings. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology exhibits characteristics that are ideal for the development of a rapid, cost-effective nucleic acid-based test for detection of HIV DNA and RNA. In this study, a sequence-specific detection method was developed for immediate, naked-eye visualization of RT-LAMP products with high sensitivity and specificity. The rapid detection method was incorporated into the HIV-1-specific RT-LAMP assay and validated using minute volumes of whole blood from HIV-1-infected individuals. Together with the minimal sample preparation time and one-step, isothermal amplification reaction, the sequence-specific detection method adds to the overall versatility of the RT-LAMP assay and enhances the applicability for use at point-of-care or resource-limited sites.

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Direct detection of human herpesvirus 6 DNA in serum by the loop-mediated isothermal amplification method

Cited by 53 publications

References 34 publications

Direct Detection of Equine Herpesvirus Type 1 DNA in Nasal Swabs by Loop-Mediated Isothermal Amplification (LAMP)

Direct Detection of Equine Herpesvirus Type 1 DNA in Nasal Swabs by Loop-Mediated Isothermal Amplification (LAMP)

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye

Sequence‐specific detection method for reverse transcription, loop‐mediated isothermal amplification of HIV‐1

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