Development of SYBR Green I Based Real-Time RT-PCR Assay for Specific Detection of Watermelon silver mottle Virus

Xueqin, Rao; Sun, Jiazhi

doi:10.15171/ijb.1124

Cited by 6 publications

(6 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the preliminary study titled "Developed HER-2 I655V detection based on SYBR Green Ibased melting curve method," it was shown that an unsharp melt curve target and a clearly additional curve appear at temperatures of 75°C. Additional curve at 75°C indicates the existence of a dimer primer which means low specificity [21][22][23][24]. To develop a real-time PCR research validation, the specificity, repeatability, reproducibility, and stability parameters of the assays were considered [25,26].…”

Section: Discussionmentioning

confidence: 99%

Design and characterization of a SYBR Green I-based melting curve method for investigation of HER2I655V polymorphism in breast cancer

Desriani

Azamris

Ghaissani

et al. 2021

Journal of Genetic Engineering and Biotechnology

View full text Add to dashboard Cite

Background Breast cancer is a disease in which cell grows rapidly forming a mass in the breast. HER2 polymorphisms Ile655Val have been studied as biomarkers for breast cancer and may comprise a risk factor of cardiac toxicity for breast cancer-consuming trastuzumab. Aim of work: In this study, we developed a simple, low cost, and rapid test to detect polymorphism at HER2 gene using SYBR Green I-based melting curve method. Subjects and methods In this report, we performed allelic discrimination with real-time temperature melting (Tm) Shift SYBR Green I-based melting curve method. The melting profiles of amplified DNA HER-2 Ile655Val and its characteristics were analyzed. Result Tm value of HER2 GG and AA alleles were 85 ± 0.14 °C and 82.5 ± 0.23 °C, respectively, while cycle threshold (Ct) value of GG, AG, and AA alleles were 19.6 ± 0.27, 22.5 ± 0.23, 18.6 ± 0.22 correspondingly; furthermore, no template control has shown consisting Ct value at 31.18 ± 0.27. The developed methods’ characteristics were optimum annealing at 62 °C and Kappa coefficient value 1 with the mean almost consistent with PCR-sequencing. The coefficient of variability for intra-assay of GG, AG, and AA was in the range of 0.2–1%, while the coefficient of variability for inter-assay for each were in the range 0.7–1%. Further, based on PCR, shelf-life assay has shown stability for 3 months of storage observation. Conclusion This approach may be considered as simple, rapid, and low cost supporting the rapid study of HER2 epidemiology. Furthermore, the developed methods potentially facilitate clinicians in dealing with breast cancer patients, especially in considering about the cardiotoxicity effect of trastuzumab.

show abstract

Section: Discussionmentioning

confidence: 99%

Design and characterization of a SYBR Green I-based melting curve method for investigation of HER2I655V polymorphism in breast cancer

Desriani

Azamris

Ghaissani

et al. 2021

Journal of Genetic Engineering and Biotechnology

View full text Add to dashboard Cite

show abstract

“…In these regards immunogenicity of the multi-epitopic recombinant glycoproteins virus: implications may be concerned (48). There are also several biotechnological research on the virus (49)(50)(51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61), each of which can be helpful in applying the results. There are several limitations to this study.…”

Section: Discussionmentioning

confidence: 99%

Importance of miR-125a-5p and miR-122-5p expression in patients with HBV infection

Lak

Yaghobi

Garshasbi

2020

Cell Mol Biol (Noisy-le-grand)

View full text Add to dashboard Cite

MicroRNAs (miRNAs) as a small RNA and post-transcriptional modulator are shown to have regulatory effects for different cellular activities and pathways, such as metabolism, virus replication and also cell growth. In addition, miRNAs can regulate the replication of the hepatitis B virus (HBV). Therefore, the expression profile of miRNAs was evaluated in HBV infected patient groups and healthy controls. The expression levels of following microRNAs (as noninvasive biomarkers) were compared in two experimental (those with various stages of HBV infection) and control groups to evaluate their diagnosis ability: miR-122-5p, miR-125a-5p, miR-199a-3p, miR-210-5p, miR-205-5p, miR-155-5p, miR-372-5p, and miR-1-5p. RNA extraction was performed for 45 serum samples. The miRCURY LNA™ Universal RT-miRNA-PCR system and miRNA PCR panels were used for measuring microRNA expression profiles. To normalize quantitative values, the endogenous reference by UniSp6 expression was used.Serum miR-125a-5p and miR-122-5p were significantly higher in patients in different stages of HBV infection (p<0.001) than in controls (p<0.001). Receiver operating characteristic (ROC) curve analyses suggested that serum miR-125a-5p and miR-122-5p have significant diagnostic value for HBV infection. A significant difference was not found in terms of serum levels of other miRs (miR-199a-3p, miR-210-5p, miR-205-5p, miR-155-5p, miR-372-5p and miR-1-5p). Results suggest that miR-125a-5p and miR-122-5p can be used as possible noninvasive biomarkers for monitoring of HBV infection need to confirm in future completed studies.

show abstract

“…Two main qPCR based quantification techniques, relative and absolute, have emerged from studies on virus detection. Absolute quantification with a known standard is the most commonly used for plant virus quantification (Herrera-vásquez et al, 2015;Rao and Sun, 2015;Shafiq et al, 2017). On the other hand, relative quantification uses a comparative quantification with internal reference endogenous housekeeping control genes (Bester et al, 2014;Kundu, 2012;Liu et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…Quantitative PCR (qPCR) has been used to detect and measure nucleic acids of a wide range of plant viruses with RNA or DNA genomes (Ammara et al, 2017;Bester et al, 2014;Rao and Sun, 2015;Shafiq et al, 2017). Recently a betasatellite has been reported as a key regulator of symptom induction.…”

Section: Introductionmentioning

confidence: 99%

Detection and absolute quantification of betasatellites associated with okra yellow vein mosaic disease by qPCR

Tharmila

Emmanuel

Devika

et al. 2020

Journal of Virological Methods

View full text Add to dashboard Cite

Okra yellow vein mosaic disease (OYVMD) causes serious loss in okra production in Sri Lanka. Therefore, screening of resistant okra verities is an essential need to control the disease. As the available qualitative and semi-quantitative methods failed to detect latent infection the present study aimed to develop a quantitative PCR (qPCR) assay to detect and quantify one of the OYVMD causing agent, symptom modulating satellite molecules. A pair of primers targeting a portion of βC1 gene of BYVMBs was designed and used to quantify of BYVMBs by absolute quantification method using SYBR Green I chemistry. Standard curves

show abstract

Development of SYBR Green I Based Real-Time RT-PCR Assay for Specific Detection of Watermelon silver mottle Virus

Cited by 6 publications

References 25 publications

Design and characterization of a SYBR Green I-based melting curve method for investigation of HER2I655V polymorphism in breast cancer

Design and characterization of a SYBR Green I-based melting curve method for investigation of HER2I655V polymorphism in breast cancer

Importance of miR-125a-5p and miR-122-5p expression in patients with HBV infection

Detection and absolute quantification of betasatellites associated with okra yellow vein mosaic disease by qPCR

Contact Info

Product

Resources

About