DNA Lateral Flow Assay for LDLR Mutation in Familial Hypercholesterolemia

| Posted in: Science

Journal name: The Malaysian Journal of Medical Sciences
Original article title: Development of DNA-Based Lateral Flow Assay for Detection of LDLR Gene Mutation for Familial Hypercholesterolemia
The Malaysian Journal of Medical Sciences (MJMS) is a peer-reviewed, open-access journal published online at least six times a year. It covers all aspects of medical sciences and prioritizes high-quality research.
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Original source:

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Author:

Lina Khalida Saidi, Zam Zureena Md Rani, Siti Aishah Sulaiman, Rahman Jamal, Aziah Ismail, Anis Amirah Alim, Sharipah Nadzirah Syed Ahmad Ayob, Chang Fu Dee, Azrul Azlan Hamzah, Nor Azian Abdul Murad


The Malaysian Journal of Medical Sciences:

(A peer-reviewed, open-access journal)

Full text available for: Development of DNA-Based Lateral Flow Assay for Detection of LDLR Gene Mutation for Familial Hypercholesterolemia

Year: 2024 | Doi: 10.21315/mjms2024.31.3.6

Copyright (license): CC BY 4.0


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Summary of article contents:

Introduction

Genotyping techniques aimed at detecting single nucleotide polymorphisms (SNPs) often involve complex procedures and expensive equipment, limiting their accessibility in many laboratories. To address this issue, a DNA-based lateral flow assay (LFA) was developed as a point-of-care diagnostic tool for genotyping. This study focused on the specific E101K variant in the low-density lipoprotein receptor (LDLR) gene, which is associated with familial hypercholesterolemia (FH). By utilizing a more straightforward and rapid testing method, the study seeks to facilitate better access to genetic testing for individuals at risk of genetic conditions.

Summary of One Important Concept: Sensitivity and Specificity of the LFA

In the context of this study, the LFA showed impressive diagnostic efficacy with a sensitivity and specificity of 100% regarding the detection of the E101K variant. The study involved 103 hypercholesterolemic individuals, of which 5 (4.8%) tested positive for the E101K variant, consistent with the results obtained through both Sanger sequencing and the Agena MassARRAY® technique. This high sensitivity and specificity indicate that the developed LFA is a reliable tool for detecting specific genetic mutations, which is critical for accurate diagnosis and subsequent management of familial hypercholesterolemia, allowing for early intervention strategies.

Conclusion

The research highlights the potential of the LFA as an effective, accessible, and cost-efficient diagnostic tool for detecting the E101K variant in the LDLR gene. The findings demonstrate the assay’s applicability for supporting genetic screenings, potentially screening family members of affected individuals, thus facilitating timely medical intervention. The LFA's success not only signifies advancements in point-of-care genetic testing but also emphasizes its role in broadening the accessibility and practicality of genetic diagnostics in clinical settings, ultimately enhancing patient care for conditions linked to genetic mutations.

FAQ section (important questions/answers):

What is the main purpose of the study?

The study aimed to develop a deoxyribonucleic acid (DNA)-based lateral flow assay (LFA) as a point-of-care test (POCT) diagnostic tool for genotyping a specific single nucleotide variation (E101K) associated with familial hypercholesterolemia.

How many individuals tested positive for the E101K variant?

Out of 103 hypercholesterolemic individuals, 5 (approximately 4.8%) tested positive for the E101K variant in the LDLR gene, confirming their diagnosis of definite familial hypercholesterolemia through subsequent sequencing.

What were the sensitivity and specificity results for LFA detection?

The sensitivity and specificity of the LFA detection method for the E101K variant were both found to be 100% when compared to results obtained using the Agena MassARRAY® genotyping method.

What are the potential applications of the developed LFA?

The developed LFA can be used in point-of-care settings for detecting the E101K variant. It can also be utilized for screening family members potentially affected by familial hypercholesterolemia and for detecting other single nucleotide polymorphisms.

Glossary definitions and references:

Scientific and Ayurvedic Glossary list for “DNA Lateral Flow Assay for LDLR Mutation in Familial Hypercholesterolemia”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.

1) Table:
In scientific literature, a table presents data in an organized manner, facilitating comparison and analysis. Tables often summarize results, demographics, or methodological details succinctly, making it easier for readers to grasp complex information quickly. They serve as visual aids, enhancing the clarity of findings in a study.

2) Mutation:
A mutation is a change in the DNA sequence that can affect an organism's traits. Mutations can occur naturally or be induced by environmental factors. They can lead to genetic variation, some of which may result in diseases, while others might confer beneficial traits, playing a significant role in evolution.

3) Study (Studying):
In academic and scientific contexts, a study refers to a detailed investigation or analysis of a particular subject or phenomenon. It involves systematic collection of data and evidence to draw conclusions. Studies can contribute to the body of knowledge in a field, support theories, or influence practice.

4) Line:
In biological and genetic studies, a line often refers to a lineage or sequence, such as those seen in genetic testing results. The term is also used in diagnostic tests to describe the visual indicators of results, where the presence of a line indicates a positive response in assays.

5) Disease:
A disease is a pathological condition of a bodily part, characterized by an abnormal functioning or structure, often resulting in specific symptoms. Diseases can be caused by various factors, including genetic mutations, infections, environmental influences, and lifestyle choices, and can significantly impact public health and individual well-being.

6) Gold (Golden):
In the context of lateral flow assays (LFAs), gold nanoparticles are commonly used as labels due to their unique properties, such as high stability and strong optical signals. These nanoparticles enhance the visibility of results, aiding in the detection of target analytes through colorimetric changes in assays.

7) Family:
In genetics, family can refer to a group of related individuals genetically linked by inheritance. Understanding hereditary patterns within families is crucial in studying genetic diseases, such as familial hypercholesterolemia, as it helps identify at-risk individuals and inform appropriate screening and management strategies.

8) Viru:
While 'viru' itself may not be a complete word, it is often linked to 'virus', referring to infectious agents that can cause diseases in hosts. Understanding viral infections is critical for public health, especially in the context of diseases that have epidemic potential or require specific diagnostic techniques.

9) Performance:
Performance in a scientific context usually refers to the effectiveness and efficiency of a method or assay. It encompasses measures such as sensitivity, specificity, accuracy, and reliability, which are critical for determining how well a diagnostic tool operates in detecting or quantifying a particular analyte.

10) Developing:
The term developing is often associated with the process of creating or improving methods, technologies, or products. In biomedical research, developing new diagnostic tools, such as lateral flow assays or genetic tests, is essential for enhancing disease detection, improving public health responses, and providing better patient care.

11) Science (Scientific):
Scientific pertains to the methods and principles of science that involve systematic observation, experimentation, and analysis. Emphasizing scientific rigor and reproducibility is essential in research to ensure findings are valid and contribute to the body of knowledge effectively.

12) Purification:
In molecular biology, purification refers to the processes employed to isolate specific molecules or compounds from a mixture. This is integral in developing assays and diagnostic tools where the specificity and accuracy of detection depend on the removal of contaminants, ensuring that only the desired analyte is measured.

13) Calculation:
Calculation in scientific studies often involves statistical methods to analyze data and determine significant outcomes. It is essential for determining sample sizes, validating results, and interpreting performance metrics in diagnostic tests. Accurate calculations are critical for ensuring the robustness and reliability of scientific findings.

14) Epidemic:
An epidemic refers to an outbreak of a disease that occurs suddenly and spreads rapidly among a population. Understanding the dynamics of epidemics is crucial for public health planning, response strategies, and developing vaccines or treatments, especially in cases involving infectious diseases that can have widespread impacts.

15) Rules:
In scientific research, rules can refer to the established guidelines or protocols that ensure the validity and reliability of experiments and studies. Adhering to these rules is essential for maintaining ethical standards, obtaining accurate results, and ensuring that research findings can be reproduced by others.

16) Blood:
Blood is a vital fluid in vertebrates that delivers essential nutrients and oxygen to the body while removing waste products. In genetic studies, blood samples are often used to extract DNA for analysis, enabling the identification of genetic disorders and the study of familial diseases and mutations.

17) Cina:
China may refer to the geographical and cultural context in which certain studies are conducted or may indicate the origin of specific genetic variants in populations. Knowledge of population genetics in regions like China can enhance understanding of the distribution and prevalence of genetic diseases globally.

18) Accumulation (Accumulating, Accumulate):
Accumulation refers to the gradual gathering or increase of a substance over time. In a biological context, it can describe the build-up of certain compounds, such as lipids or toxins, within an organism, which can lead to health issues or inform research regarding genetic conditions or environmental health.

19) Thalassemia:
Thalassemia is a genetic blood disorder characterized by reduced hemoglobin production, leading to anemia. Genetic testing for thalassemia typically involves identifying mutations in the globin genes. Understanding this condition is crucial for early diagnosis, management, and providing genetic counseling to affected families.

20) Fixation:
In biological research, fixation is a process used to preserve cells or tissue samples by stabilizing their structures. This is essential for microscopic examination and various assays, allowing researchers to study cellular components and molecular markers accurately in the context of disease pathology or genetic studies.

21) Water:
Water is essential for life and is involved in numerous biological processes. In laboratory settings, the quality of water used for experiments and assays is crucial; impurities in water can affect the results of biochemical reactions and assays, potentially leading to inaccurate findings.

22) Arrow:
In scientific diagrams or graphical representations, arrows are used to indicate direction, flow, or relationships between different components. In assays, arrows can signify the movement of fluid through different parts of a lateral flow device or illustrate pathways in biochemical processes, aiding comprehension.

23) Fever:
Fever is a common physiological response to infection, characterized by an elevated body temperature. In the context of public health, monitoring fever is essential for diagnosing infectious diseases. Understanding fever mechanisms can inform treatment strategies and epidemiological studies, especially during epidemic outbreak investigations.

24) Nara:
Nara may refer to a place or could be a part of specific studies or experiments involving geographical or genetic contexts. The significance of studying different regions, such as Nara, adds to the understanding of genetic diversity or disease prevalence patterns across populations.

25) Hand:
In research, the term 'hand' might not have a specific scientific meaning but can refer to human involvement in laboratory processes or the handling of specimens and equipment. Proper technique in handling samples is critical to ensure valid and reliable results in scientific studies.

26) Life:
Life refers to the condition that distinguishes organisms from inorganic matter, encompassing the biological processes of growth, reproduction, and response to stimuli. Understanding life at molecular and genetic levels is crucial for advancements in medicine, genetics, and various biological sciences.

27) Rheumatoid arthritis:
Rheumatoid arthritis is an autoimmune disorder characterized by chronic inflammation of joints, leading to pain and disability. Understanding its genetic factors is essential for developing targeted therapies and improving diagnostic methods, as early detection can significantly enhance patient outcomes and quality of life.

28) Discussion:
In scientific literature, the discussion section contextualizes research findings, interprets results, and compares them with existing knowledge. It is crucial for addressing implications, limitations, and potential future studies. Effective discussion enhances the impact and understanding of the research within the broader scientific community.

29) Relative:
Relative often refers to comparison with something else in a scientific context. It can describe genetic relationships among organisms, the frequency of traits, or comparisons between different study results, aiding in understanding genetic diversity, inheritance patterns, or the significance of findings.

30) Cutting:
In research, cutting may refer to the act of slicing biological specimens for analysis or in genetic contexts referring to cutting DNA strands using restriction enzymes during cloning and molecular manipulation. Proper techniques in cutting are crucial for ensuring sample integrity and achieving accurate results.

31) Account:
In a scientific context, an account may refer to a detailed explanation or record of research findings or methodologies. Maintaining accurate accounts of procedures and results is essential for reproducibility, transparency in research practices, and for advancing collective knowledge in scientific fields.

32) Humana:
Humana typically refers to the publishing company Humana Press, known for publishing academic and scientific research. Materials published by such organizations often contribute significantly to various scientific domains, providing valuable information, methodologies, and insights for researchers and practitioners.

33) Medium:
In microbiology and cell culture, a medium refers to the nutrient-rich solution used to support the growth of cells or microorganisms. The composition of the medium can significantly influence experimental outcomes, making its selection critical for successful laboratory experiments.

34) Glass:
Glass is a commonly used material in laboratories, especially for containers such as test tubes and flasks. Its chemical resistance and transparency make it ideal for various experiments. Understanding the properties of glassware is essential in laboratory practices to ensure accurate results in experiments.

35) Nagel:
Nagel often refers to Macherey-Nagel, a company known for producing laboratory equipment and supplies, particularly in molecular biology and nucleic acid applications. Utilizing products from reputable suppliers like Nagel ensures reliability and quality in research methodologies and experiments.

36) King:
King may refer to ThermoFisher's KingFisher Duo Prime, a machine used for DNA extraction. This equipment simplifies sample preparation steps, enhancing laboratory efficiency and is significant in genetic studies where high-quality DNA is essential for valid experiments.

37) Rich (Rch):
In a scientific context, the term 'rich' often describes environments or media with a high concentration of nutrients or components that promote growth or activity, particularly in microbiology or cell cultures. Rich media are crucial for optimizing conditions for experimentation and research.

38) Post:
Post may refer to various contexts in research, often indicating after a certain event, such as post-treatment outcomes or post-experimental analyses. Clarifying the timing of observations and analyses is critical for understanding the effects and significance of experimental interventions.

39) Drug:
A drug is a substance used to diagnose, cure, treat, or prevent disease. Understanding the genetic factors influencing drug metabolism and responses is vital in pharmacogenomics to optimize treatment plans and reduce adverse effects in patients, enhancing personalized medicine approaches.

40) Male:
In biological and medical studies, 'male' refers to the sex of an organism that typically produces sperm and has specific genetic or physiological characteristics. Investigating male-specific traits or diseases is crucial in understanding sex-based differences in health outcomes and disease prevalence.

Other Science Concepts:

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Discover the significance of concepts within the article: ‘DNA Lateral Flow Assay for LDLR Mutation in Familial Hypercholesterolemia’. Further sources in the context of Science might help you critically compare this page with similair documents:

Doi, Medical history, Statistical analysis, Sample size, Dyslipidaemia, Demographic Data, LDL cholesterol, Atherosclerosis, Polymerase chain reaction, Lipid profile, Chi square test, COVID 19, Diagnostic tool, Single nucleotide polymorphism, PMC free article, PubMed, Google Scholar, Sample preparation, Genetic testing, DNA Analysis, Agarose gel electrophoresis, PCR amplification, Bioinformatics analysis, Gel electrophoresis, Sensitivity and Specificity, Ethanol precipitation, Wild type, PCR analysis, DNA extraction, Sequence analysis, DNA sequencing, Next generation sequencing, PCR product, Statin treatment, PCR assay, Genetic screening, Point of care diagnostics, Nucleic Acid Detection, Gene variants, Low-density lipoprotein receptor, Gold nanoparticle, Whole-exome sequencing, Sanger sequencing, Control sample, Familial Hypercholesterolemia, Exome sequencing, Capillary action, Forward primer, Reverse primer, PCR process, DNA Mutations, Point-of-care test, Genotyping method, Microarray, Colloidal Gold Nanoparticles, Clinical practice guideline, PCR amplicon, J.Lipid Res, MALDI-TOF-MS, PCR-based assays, Primer design, Int J Mol Sci, PLoS ONE, Result interpretation, Hypoalphalipoproteinemia.

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