scholarly journals A Novel Isothermal Amplification Method for Detecting Mouse Source Component in Meat

2019 ◽  
Vol 102 (3) ◽  
pp. 872-877 ◽  
Author(s):  
Xinnan Li ◽  
Xueqin Gao ◽  
Yifu Guan
Keyword(s):  
Isothermal Amplification ◽  
New Method ◽  
Simple Method ◽  
New Approach ◽  
Safety Issues ◽  
Amplification Method ◽  
Specificity And Sensitivity ◽  
Great Specificity ◽  
Nonspecific Amplification ◽  
Dna Template

Abstract Background: The problem of adulterated meat has become one of the greatest food safety issues in the world. It is reported that the meat used for adulteration includes fox meat, raccoon meat, mink meat, mouse meat, and so on. Although this kind of meat is edible in some areas, such meat is potentially harmful to human health because it is easy to carry bacteria, viruses, and harmful substances. The harm of mouse meat is most frightening. Therefore, it is urgent to develop a fast, accurate, and simple method to effectively identify mouse meat. Methods: In the present study, a new method of isothermal amplification based on the 16S ribosomal RNA gene of the mitochondrial DNA of the mouse was developed. The method is meant to improve the loop-mediated isothermal amplification (LAMP), separating the forward inner primers and backward inner primers, greatly reducing the nonspecific amplification of the method. Results: We have successfully obtained a set of best primers. The developed system allowed for the detection of 0.5% mouse meat from meat mixture effectively and specifically. The best ratio of the primers (F3: F2: F1: RF) was 1:2:2:8, and the optimum concentration of DNA template was 0.35 ng/μL. Conclusions: The assay has great specificity and sensitivity for detecting mouse meat and could provide specific positive results within 1 h. Highlights: We found a new approach of isothermal amplification to detect mouse source components. The LOD is determined to be 0.5 mg/mg. This new method is easy to perform and is able to provide rapid results in the specific detection of mouse meat sources.

scholarly journals Detection and discrimination of multiple strains of Zika virus by reverse transcription-loop-mediated isothermal amplification

2020 ◽  
Vol 48 (1) ◽  
Author(s):  
Hiroka Aonuma ◽  
Itoe Iizuka-Shiota ◽  
Tokio Hoshina ◽  
Shigeru Tajima ◽  
Fumihiro Kato ◽  
...  
Keyword(s):  
Virus Strain ◽  
Reverse Transcription ◽  
Zika Virus ◽  
Virus Disease ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Simple Method ◽  
Loop Mediated Isothermal Amplification ◽  
Specificity And Sensitivity ◽  
Virus Strains

Abstract Background Monitoring both invasion of Zika virus disease into free countries and circulation in endemic countries is essential to avoid a global pandemic. However, the difficulty lies in detecting Zika virus due to the large variety of mutations in its genomic sequence. To develop a rapid and simple method with high accuracy, reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was adopted for the detection of Zika virus strains derived from several countries. Results Common primers for RT-LAMP were designed based on the genomic sequences of two standard Zika strains: African lineage, MR-766, and Asian lineage, PRVABC59. RT-LAMP reactions using a screened primer set, targeting the NS3 region, detected both Zika virus strains. The minimum detectable quantity was 3 × 10−2 ng of virus RNA. Measurable lag of reaction times among strains was observed. The RT-LAMP method amplified the target virus sequence from the urine and serum of a patient with a travel history in the Caribbean Islands and also provided a prediction about which lineage of Zika virus strain was present. Conclusions The RT-LAMP method using a well-optimized primer set demonstrated high specificity and sensitivity for the detection of Zika virus strains with a variety in genomic RNA sequences. In combination with the simplicity of LAMP reaction in isothermal conditions, the optimized primer set established in this study may facilitate rapid and accurate diagnosis of Zika fever patients with virus strain information.

scholarly journals Microfluidic Chip with Two-Stage Isothermal Amplification Method for Highly Sensitive Parallel Detection of SARS-CoV-2 and Measles Virus

Micromachines ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1582
Author(s):  
Qin Huang ◽  
Xiaohui Shan ◽  
Ranran Cao ◽  
Xiangyu Jin ◽  
Xue Lin ◽  
...  
Keyword(s):  
Microfluidic Chip ◽  
Measles Virus ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Nasopharyngeal Swab ◽  
Two Stage ◽  
Parallel Detection ◽  
Amplification Method ◽  
Specificity And Sensitivity

A two-stage isothermal amplification method, which consists of a first-stage basic recombinase polymerase amplification (RPA) and a second-stage fluorescence loop-mediated isothermal amplification (LAMP), as well as a microfluidic-chip-based portable system, were developed in this study; these enabled parallel detection of multiplex targets in real time in around one hour, with high sensitivity and specificity, without cross-contamination. The consumption of the sample and the reagent was 2.1 μL and 10.6 μL per reaction for RPA and LAMP, respectively. The lowest detection limit (LOD) was about 10 copies. The clinical amplification of about 40 nasopharyngeal swab samples, containing 17 SARS-CoV-2 (severe acute respiratory syndrome coronavirus) and 23 measles viruses (MV), were parallel tested by using the microfluidic chip. Both clinical specificity and sensitivity were 100% for MV, and the clinical specificity and sensitivity were 94.12% and 95.83% for SARS-CoV-2, respectively. This two-stage isothermal amplification method based on the microfluidic chip format offers a convenient, clinically parallel molecular diagnostic method, which can identify different nucleic acid samples simultaneously and in a timely manner, and with a low cost of the reaction reagent. It is especially suitable for resource-limited areas and point-of-care testing (POCT).

A New Approach for the Analysis of Mixture Toxicity Data

1992 ◽  
Vol 26 (9-11) ◽  
pp. 2345-2348 ◽  
Author(s):  
C. N. Haas
Keyword(s):  
Quantitative Analysis ◽  
Mixture Toxicity ◽  
New Method ◽  
Metal Exposure ◽  
Positive Interactions ◽  
Toxicity Data ◽  
Data Set ◽  
New Approach ◽  
Negative Interactions

A new method for the quantitative analysis of multiple toxicity data is described and illustrated using a data set on metal exposure to copepods. Positive interactions are observed for Ni-Pb and Pb-Cr, with weak negative interactions observed for Ni-Cr.

scholarly journals New Method for Generating New Families of Distributions

Symmetry ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 726
Author(s):  
Lamya A. Baharith ◽  
Wedad H. Aljuhani
Keyword(s):  
Exponential Distribution ◽  
Real Data ◽  
Rate Function ◽  
New Method ◽  
Likelihood Method ◽  
Alpha Power ◽  
Unknown Parameters ◽  
Failure Rate Function ◽  
New Approach ◽  
Numerical Studies

This article presents a new method for generating distributions. This method combines two techniques—the transformed—transformer and alpha power transformation approaches—allowing for tremendous flexibility in the resulting distributions. The new approach is applied to introduce the alpha power Weibull—exponential distribution. The density of this distribution can take asymmetric and near-symmetric shapes. Various asymmetric shapes, such as decreasing, increasing, L-shaped, near-symmetrical, and right-skewed shapes, are observed for the related failure rate function, making it more tractable for many modeling applications. Some significant mathematical features of the suggested distribution are determined. Estimates of the unknown parameters of the proposed distribution are obtained using the maximum likelihood method. Furthermore, some numerical studies were carried out, in order to evaluate the estimation performance. Three practical datasets are considered to analyze the usefulness and flexibility of the introduced distribution. The proposed alpha power Weibull–exponential distribution can outperform other well-known distributions, showing its great adaptability in the context of real data analysis.

Identification for adulteration of beef with chicken based on single primer-triggered isothermal amplification

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Jiao Chen ◽  
Pansong Zhang ◽  
Haixia Wang ◽  
Yanjing Shi
Keyword(s):  
Limit Of Detection ◽  
Chicken Meat ◽  
Isothermal Amplification ◽  
Work Flow ◽  
Acid Extraction ◽  
Specificity And Sensitivity ◽  
Total Dna ◽  
Meat Species ◽  
Species Specific ◽  
Single Primer

Abstract Adulteration of beef with cheap chicken has become a growing problem worldwide. In this study, a quick, single primer-triggered isothermal amplification (SAMP) combined with a fast nucleic acid extraction method was employed to detect the chicken meat in adulterated beef. Chicken from adulterated beef was identified using the chicken species-specific primer designed according to the Gallus gallus mitochondrial conserved sequences. Our SAMP method displayed good specificity and sensitivity in detecting chicken and beef meat DNA–the limit of detection (LOD) of SAMP is 0.33 pg/μL of chicken and beef total DNA and 2% w/w chicken meat in beef. The whole work flow from DNA extraction to signal detection can be finished within 1 h, fulfilling the requirement of on-site meat species identification.

Capturing Protein Droplets: Label-free Visualization and Detection of Protein Liquid-Liquid Phase Separation with an Aggregation-Induced Emission Fluorogen

2021 ◽  
Author(s):  
Yan-Mei Li ◽  
Chu-Qiao Liang ◽  
Lin Wang ◽  
Yun-Yi Luo ◽  
Qian-Qian Li
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Liquid Phase Separation ◽  
New Method ◽  
Label Free ◽  
Simple Method ◽  
Aggregation Induced Emission ◽  
Liquid Liquid Phase Separation

We developed a new method for protein droplet visualization by means of a droplet probe (DroProbe) based on an aggregation-induced emission (AIE) fluorogen. A simple method for viscosity comparison of...

scholarly journals P02.06 A Novel Loop-Mediated Isothermal Amplification Method for Robust Detection of EGFR Mutations

2021 ◽  
Vol 16 (3) ◽  
pp. S248-S249
Author(s):  
Y. Saito ◽  
A. Matsui ◽  
S. Michiyuki ◽  
Y. Yamauchi ◽  
N. Takahashi ◽  
...  
Keyword(s):  
Isothermal Amplification ◽  
Egfr Mutations ◽  
Robust Detection ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method

Comparison of New Telescopic Video Microlaryngoscopic and Standard Microlaryngoscopic Techniques

1992 ◽  
Vol 101 (1) ◽  
pp. 51-60 ◽  
Author(s):  
Eiji Yanagisawa ◽  
Ken Yanagisawa ◽  
Jay B. Horowitz ◽  
Lawrence J. Mambrino
Keyword(s):  
Video Camera ◽  
New Method ◽  
Depth Of Field ◽  
Microscopic Technique ◽  
New Approach ◽  
Advantages And Disadvantages ◽  
Laryngeal Surgery ◽  
Valuable Method ◽  
Microlaryngeal Surgery ◽  
Television Image

A new approach to microlaryngeal surgery using a specially designed video microlaryngoscope with a rigid endoscopic telescope and an attached video camera was introduced by Kantor et al in 1990. The ability to video document and perform surgery of the larynx by viewing a high-resolution television image was demonstrated. This method was recommended over the standard microscopic technique for increased visibility with greater depth of field, unimpeded instrument access, instant documentation, and superior teaching value. The authors tried this new method and the standard microscopic technique at the same sitting on a series of patients. This paper will compare these two different techniques and discuss their advantages and disadvantages. Although the new method has many advantages, the standard microscopic technique remains as a valuable method in laryngeal surgery.

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