Codeposition of dNTPs Detection for Rapid LAMP‐based Sexing of Bovine Embryos

Zhang, Z. P.; Zhang, Y.; Liu, J. P.; Zhang, J. T.; An, Zhixing; Quan, F. S.; Zhang, L.; Cai, Xiaoming; Pu, Shi

doi:10.1111/j.1439-0531.2007.01006.x

Cited by 12 publications

(4 citation statements)

References 25 publications

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“…Over the past decades, it was further developed in combination with other molecular methods, including reverse transcription and multiplex amplification [39,40]. Currently, the main varieties of LAMP assay available are designed by colorimetric change [41,42], real-time monitored [39] with fluorescent dye or using turbidity meter [43]. Nowadays, LAMP is widely applied as new diagnostics for infectious diseases.…”

Section: Discussionmentioning

confidence: 99%

A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus

Hu¹,

Wan²,

Mu³

et al. 2019

J Infect Dev Ctries

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Introduction: Human respiratory syncytial virus (hRSV) is a common respiratory virus closely related to respiratory tract infection (RTI). Rapid and accurate detection of hRSV is urgently needed to reduce the high morbidity and mortality due to hRSV infection. Methodology: Here, we established a highly sensitive and specific reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid detection of A and B group hRSV simultaneously. The specific primer sets for hRSV A and B groups were designed in the M and M2-2 gene, respectively. SYTO 9 was used as the fluorescent dye for real-time monitoring of the amplification of hRSV RNA without cross reaction between hRSV A and B. Results: The limit of detection (LOD) of our new method was 281.17 50% tissue culture infective doses (TCID50)/mL for hRSV A and 1.58 TCID50/mL for hRSV B. Using 90 clinical samples, a comparison to traditional RT-PCR was performed to validate this assay. The positivity rate of RT-LAMP and RT-PCR were 67.8% and 55.6%, respectively, and the positivity rate of RT-LAMP was significantly higher than RT-PCR (χ2 test, P < 0.01). Conclusions: Compared with traditional RT-PCR method, the newly developed fluorescent RT-LAMP combined with well-designed primers and SYTO 9 is quite sensitive, specific, rapid and well applicable to hRSV clinical diagnosis.

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Section: Discussionmentioning

confidence: 99%

A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus

Hu¹,

Wan²,

Mu³

et al. 2019

J Infect Dev Ctries

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“…LAMP can amplify nucleic acids with high specificity, sensitivity and rapidity under isothermal conditions by using a DNA polymerase with strand displacement activity. LAMP products can be visualized with the naked eye by adding fluorescent DNA‐intercalating dyes such as ethidium bromide (Pham, Nakajima, Ohashi, & Onuma, ), SYBR® Green I (Notomi et al, ), propidium iodide (Hill et al, ), Quant‐iT™ PicoGreen® (Tomlinson, Barker, & Boonham, ) or GeneFinder™ (Almasi, Ojaghkandi, Hemmatabadi, Hamidi, & Aghaei, ), by adding metal ion indicators such as hydroxynaphthol blue (HNB) (Goto, Honda, Ogura, Nomoto, & Hanaki, ), CuSO 4 (Zhang et al, ), calcein (Tomita, Mori, Kanda, & Notomi, ) or pH indicators such as cresol red (Tanner, Zhang, & Evans, ). The reactions can be monitored using real‐time fluorescence measurements or a turbidity measuring system.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of Phytophthora nicotianae by simple DNA extraction and real‐time loop‐mediated isothermal amplification assay

Hieno

Li²,

Afandi

et al. 2019

Journal of Phytopathology

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Phytophthora nicotianae is a phytopathogenic oomycete with a wide host range and worldwide distribution. Rapid detection and diagnosis at the early stages of disease development are important for the effective control of P. nicotianae. In this study, we designed a simple and rapid loop‐mediated isothermal amplification (LAMP)‐based detection method for P. nicotianae. We tested three DNA extraction methods and selected the Kaneka Easy DNA Extraction Kit version 2, which is rapid and robust for LAMP‐based detection. The designed primers were tested using mycelial DNA from 35 species (81 isolates) of Phytophthora, 12 species (12 isolates) of Pythium, one isolate of Phytopythium and one isolate each from seven other soil‐borne pathogens. All of the 42 P. nicotianae isolates were detected by these primers, and no other isolates gave positive results. Three isolates were tested for the sensitivity of the reaction, and the lowest amounts of template DNA that could be detected were 10 fg for two isolates and 1 fg for the third. The target was detected within 25 min in all tested samples, including DNA extracted from both inoculated and naturally infected plants. In contrast, PCR assays with P. nicotianae‐specific primers failed or showed weakened detection in several samples. Thus, we found that the rapid DNA extraction and LAMP assay methods developed in this study can be used to detect P. nicotianae with high sensitivity, specificity and stability.

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“…Loop Mediated Isothermal Amplification (LAMP) is a relatively new, less complicated, more rapid DNA detection method, which has been developed as an alternative to PCR. Moreover, LAMP has been used to determine the sex of bovine embryos, female calves, and Columbidae birds [ 2 , 3 ] as well as detect viruses such as Taura syndrome in shrimp [ 4 ]. Kanchanaphum et al [ 5 ] developed and used the LAMP method to determine the sex of humans using the SRY gene as a target.…”

Section: Introductionmentioning

confidence: 99%

Time Course of Detection of Human Male DNA from Stained Blood Sample on Various Surfaces by Loop Mediated Isothermal Amplification and Polymerase Chain Reaction

Kanchanaphum

2018

BioMed Research International

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This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP), and LAMP-Lateral Flow Dipstick (LFD). For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, wood, clay, and tile. Then, the samples were stored at room temperature for 1, 7, 30, and 60 day(s). After the DNA amplification, the gel electrophoresis process was applied to detect LAMP product. The LFD was combined with the LAMP to detect LAMP product on the male cloth samples. For the male samples, the time course of detection on the first and seventh days indicated positive for both LAMP and PCR products on all the surfaces while no DNA amplification was found on any of the female samples. On day 30, positive LAMP product was still found on all the male samples. However, it had faded on the tiles. Moreover, all the male samples, which had tested positive for PCR product, were blurred and unclear. On day 60, LAMP product was still found on all the male samples. Conversely, the PCR method resulted in no bands showing for any of the male samples. However, the LAMP-LFD method detected product on all the male samples of cloth. The results show that the LAMP is an effective, practical, and reliable molecular-biological method. Moreover, the LFD can increase the efficiency and sensitivity of the LAMP, making it more suitable for field studies because gel electrophoresis apparatus is not required.

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Codeposition of dNTPs Detection for Rapid LAMP‐based Sexing of Bovine Embryos

Cited by 12 publications

References 25 publications

A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus

A quite sensitive fluorescent loop-mediated isothermal amplification for rapid detection of respiratory syncytial virus

Rapid detection of Phytophthora nicotianae by simple DNA extraction and real‐time loop‐mediated isothermal amplification assay

Time Course of Detection of Human Male DNA from Stained Blood Sample on Various Surfaces by Loop Mediated Isothermal Amplification and Polymerase Chain Reaction

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