Optimization of Analytical Procedures Reduces Variability of Results
|
By LabMedica International staff writers Posted on 16 Jul 2018 |
|
Image: Reducing variability of quantitative suspension array assays is critical to the success of multi-center and large sero-epidemiological studies (Photo courtesy of Pau Fabregat).
A team of Spanish researchers recently described the development and optimization of several "quantitative suspension array" assays (qSAT) designed to assess natural and vaccine-induced responses to malaria and other parasites.
Reducing variability of quantitative suspension array assays is critical to the success of multi-center and large sero-epidemiological studies. Towards this end, investigators at the Barcelona Institute for Global Health (Spain) analyzed the effect of various procedural conditions on the variability of an in-house IgG multiplex assay designed to detect different types and classes of antibodies against multiple P. falciparum antigens.
The assay comprised different parasite antigens coupled to microspheres. These beads were tested to determine if they were recognized by plasma samples from individuals who may have been exposed to malaria. The assay was designed to detect different types and classes of antibodies against multiple P. falciparum antigens, in a sensitive and specific manner. The simple multiplex assay was found to be highly reproducible not only between experiments but also between operators and laboratories.
The investigators identified the key factors requiring optimization to reduce variability of the assay. Then, they used the assay to better characterize the antibody profile of the anti-malaria human plasma pool provided by the WHO as reference reagent.
IgG levels against seven P. falciparum antigens with heterogeneous immunogenicities were measured in test samples, in a positive control, and in blanks. Variability associated with each combination of conditions and their interactions was assessed, and the minimum number of samples and blank replicates required to achieve good replicability was determined. Results showed that antigen immunogenicity and sample seroreactivity defined the optimal dilution for assessing the effect of assay conditions on variability. Furthermore, unique antigen-bead coupling, pre-dilution of samples, incubation at 22 degrees Celsius, and an automated washing procedure reduced variability.
"These simple and reproducible multiplex protocols will allow us to analyze in detail natural and vaccine-induced antibody responses to large panels of P. falciparum antigens, and elucidate correlates of malaria protection," said senior author Dr. Carlota Dobaño, an associate research professor at the Barcelona Institute for Global Health. "Furthermore, the assay is highly versatile and can be adapted to study responses to antigens from other microbes or vaccines. We are currently collaborating with several groups outside the malaria field."
The study was published in the July 2, 2018, online edition of the journal PLOS One.
Related Links:
Barcelona Institute for Global Health
Reducing variability of quantitative suspension array assays is critical to the success of multi-center and large sero-epidemiological studies. Towards this end, investigators at the Barcelona Institute for Global Health (Spain) analyzed the effect of various procedural conditions on the variability of an in-house IgG multiplex assay designed to detect different types and classes of antibodies against multiple P. falciparum antigens.
The assay comprised different parasite antigens coupled to microspheres. These beads were tested to determine if they were recognized by plasma samples from individuals who may have been exposed to malaria. The assay was designed to detect different types and classes of antibodies against multiple P. falciparum antigens, in a sensitive and specific manner. The simple multiplex assay was found to be highly reproducible not only between experiments but also between operators and laboratories.
The investigators identified the key factors requiring optimization to reduce variability of the assay. Then, they used the assay to better characterize the antibody profile of the anti-malaria human plasma pool provided by the WHO as reference reagent.
IgG levels against seven P. falciparum antigens with heterogeneous immunogenicities were measured in test samples, in a positive control, and in blanks. Variability associated with each combination of conditions and their interactions was assessed, and the minimum number of samples and blank replicates required to achieve good replicability was determined. Results showed that antigen immunogenicity and sample seroreactivity defined the optimal dilution for assessing the effect of assay conditions on variability. Furthermore, unique antigen-bead coupling, pre-dilution of samples, incubation at 22 degrees Celsius, and an automated washing procedure reduced variability.
"These simple and reproducible multiplex protocols will allow us to analyze in detail natural and vaccine-induced antibody responses to large panels of P. falciparum antigens, and elucidate correlates of malaria protection," said senior author Dr. Carlota Dobaño, an associate research professor at the Barcelona Institute for Global Health. "Furthermore, the assay is highly versatile and can be adapted to study responses to antigens from other microbes or vaccines. We are currently collaborating with several groups outside the malaria field."
The study was published in the July 2, 2018, online edition of the journal PLOS One.
Related Links:
Barcelona Institute for Global Health
Platinum Member
COVID-19 Rapid Test
OSOM COVID-19 Antigen Rapid Test
Complement 3 (C3) Test
GPP-100 C3 Kit
New
Gold Member
Plasma Control
Plasma Control Level 1
Latest Molecular Diagnostics News
- Blood Proteins Could Warn of Cancer Seven Years before Diagnosis
- New DNA Origami Technique to Advance Disease Diagnosis
- Ultrasound-Aided Blood Testing Detects Cancer Biomarkers from Cells
- New Respiratory Syndromic Testing Panel Provides Fast and Accurate Results
- New Synthetic Biomarker Technology Differentiates Between Prior Zika and Dengue Infections
- Novel Biomarkers to Improve Diagnosis of Renal Cell Carcinoma Subtypes
- RNA-Powered Molecular Test to Help Combat Early-Age Onset Colorectal Cancer
- Advanced Blood Test to Spot Alzheimer's Before Progression to Dementia
- Multi-Omic Noninvasive Urine-Based DNA Test to Improve Bladder Cancer Detection
- First of Its Kind NGS Assay for Precise Detection of BCR::ABL1 Fusion Gene to Enable Personalized Leukemia Treatment
- Urine Test to Revolutionize Lyme Disease Testing
- Simple Blood Test Could Enable First Quantitative Assessments for Future Cerebrovascular Disease
- New Genetic Testing Procedure Combined With Ultrasound Detects High Cardiovascular Risk
- Blood Samples Enhance B-Cell Lymphoma Diagnostics and Prognosis
- Blood Test Predicts Knee Osteoarthritis Eight Years Before Signs Appears On X-Rays
- Blood Test Accurately Predicts Lung Cancer Risk and Reduces Need for Scans
Channels
Clinical Chemistry
view channel
3D Printed Point-Of-Care Mass Spectrometer Outperforms State-Of-The-Art Models
Mass spectrometry is a precise technique for identifying the chemical components of a sample and has significant potential for monitoring chronic illness health states, such as measuring hormone levels... Read more.jpg "Image: Reaching speeds up to 6,000 RPM, this centrifuge forms the basis for a new type of inexpensive, POC biomedical test (Photo courtesy of Duke University)")
POC Biomedical Test Spins Water Droplet Using Sound Waves for Cancer Detection
Exosomes, tiny cellular bioparticles carrying a specific set of proteins, lipids, and genetic materials, play a crucial role in cell communication and hold promise for non-invasive diagnostics.... Read more
Highly Reliable Cell-Based Assay Enables Accurate Diagnosis of Endocrine Diseases
The conventional methods for measuring free cortisol, the body's stress hormone, from blood or saliva are quite demanding and require sample processing. The most common method, therefore, involves collecting... Read more
Hematology
view channel
Next Generation Instrument Screens for Hemoglobin Disorders in Newborns
Hemoglobinopathies, the most widespread inherited conditions globally, affect about 7% of the population as carriers, with 2.7% of newborns being born with these conditions. The spectrum of clinical manifestations... Read more
First 4-in-1 Nucleic Acid Test for Arbovirus Screening to Reduce Risk of Transfusion-Transmitted Infections
Arboviruses represent an emerging global health threat, exacerbated by climate change and increased international travel that is facilitating their spread across new regions. Chikungunya, dengue, West... Read more
POC Finger-Prick Blood Test Determines Risk of Neutropenic Sepsis in Patients Undergoing Chemotherapy
Neutropenia, a decrease in neutrophils (a type of white blood cell crucial for fighting infections), is a frequent side effect of certain cancer treatments. This condition elevates the risk of infections,... Read more
First Affordable and Rapid Test for Beta Thalassemia Demonstrates 99% Diagnostic Accuracy
Hemoglobin disorders rank as some of the most prevalent monogenic diseases globally. Among various hemoglobin disorders, beta thalassemia, a hereditary blood disorder, affects about 1.5% of the world's... Read moreImmunology
view channel.jpg "Image: The AI predictive model identifies the most potent cancer killing immune cells for use in immunotherapies (Photo courtesy of Shutterstock)")
AI Predicts Tumor-Killing Cells with High Accuracy
Cellular immunotherapy involves extracting immune cells from a patient's tumor, potentially enhancing their cancer-fighting capabilities through engineering, and then expanding and reintroducing them into the body.... Read more
Diagnostic Blood Test for Cellular Rejection after Organ Transplant Could Replace Surgical Biopsies
Transplanted organs constantly face the risk of being rejected by the recipient's immune system which differentiates self from non-self using T cells and B cells. T cells are commonly associated with acute... Read more
AI Tool Precisely Matches Cancer Drugs to Patients Using Information from Each Tumor Cell
Current strategies for matching cancer patients with specific treatments often depend on bulk sequencing of tumor DNA and RNA, which provides an average profile from all cells within a tumor sample.... Read more
Genetic Testing Combined With Personalized Drug Screening On Tumor Samples to Revolutionize Cancer Treatment
Cancer treatment typically adheres to a standard of care—established, statistically validated regimens that are effective for the majority of patients. However, the disease’s inherent variability means... Read moreMicrobiology
view channel
Integrated Solution Ushers New Era of Automated Tuberculosis Testing
Tuberculosis (TB) is responsible for 1.3 million deaths every year, positioning it as one of the top killers globally due to a single infectious agent. In 2022, around 10.6 million people were diagnosed... Read more
Automated Sepsis Test System Enables Rapid Diagnosis for Patients with Severe Bloodstream Infections
Sepsis affects up to 50 million people globally each year, with bacteraemia, formerly known as blood poisoning, being a major cause. In the United States alone, approximately two million individuals are... Read more
Enhanced Rapid Syndromic Molecular Diagnostic Solution Detects Broad Range of Infectious Diseases
GenMark Diagnostics (Carlsbad, CA, USA), a member of the Roche Group (Basel, Switzerland), has rebranded its ePlex® system as the cobas eplex system. This rebranding under the globally renowned cobas name... Read more
Clinical Decision Support Software a Game-Changer in Antimicrobial Resistance Battle
Antimicrobial resistance (AMR) is a serious global public health concern that claims millions of lives every year. It primarily results from the inappropriate and excessive use of antibiotics, which reduces... Read morePathology
view channel
Hyperspectral Dark-Field Microscopy Enables Rapid and Accurate Identification of Cancerous Tissues
Breast cancer remains a major cause of cancer-related mortality among women. Breast-conserving surgery (BCS), also known as lumpectomy, is the removal of the cancerous lump and a small margin of surrounding tissue.... Read more
AI Advancements Enable Leap into 3D Pathology
Human tissue is complex, intricate, and naturally three-dimensional. However, the thin two-dimensional tissue slices commonly used by pathologists to diagnose diseases provide only a limited view of the... Read more
New Blood Test Device Modeled on Leeches to Help Diagnose Malaria
Many individuals have a fear of needles, making the experience of having blood drawn from their arm particularly distressing. An alternative method involves taking blood from the fingertip or earlobe,... Read more
Robotic Blood Drawing Device to Revolutionize Sample Collection for Diagnostic Testing
Blood drawing is performed billions of times each year worldwide, playing a critical role in diagnostic procedures. Despite its importance, clinical laboratories are dealing with significant staff shortages,... Read moreTechnology
view channel
New Diagnostic System Achieves PCR Testing Accuracy
While PCR tests are the gold standard of accuracy for virology testing, they come with limitations such as complexity, the need for skilled lab operators, and longer result times. They also require complex... Read more
DNA Biosensor Enables Early Diagnosis of Cervical Cancer
Molybdenum disulfide (MoS2), recognized for its potential to form two-dimensional nanosheets like graphene, is a material that's increasingly catching the eye of the scientific community.... Read more
Self-Heating Microfluidic Devices Can Detect Diseases in Tiny Blood or Fluid Samples
Microfluidics, which are miniature devices that control the flow of liquids and facilitate chemical reactions, play a key role in disease detection from small samples of blood or other fluids.... Read more
Breakthrough in Diagnostic Technology Could Make On-The-Spot Testing Widely Accessible
Home testing gained significant importance during the COVID-19 pandemic, yet the availability of rapid tests is limited, and most of them can only drive one liquid across the strip, leading to continued... Read moreIndustry
view channel
Danaher and Johns Hopkins University Collaborate to Improve Neurological Diagnosis
Unlike severe traumatic brain injury (TBI), mild TBI often does not show clear correlations with abnormalities detected through head computed tomography (CT) scans. Consequently, there is a pressing need... Read more
Beckman Coulter and MeMed Expand Host Immune Response Diagnostics Partnership
Beckman Coulter Diagnostics (Brea, CA, USA) and MeMed BV (Haifa, Israel) have expanded their host immune response diagnostics partnership. Beckman Coulter is now an authorized distributor of the MeMed... Read more_1.jpg "Image: Thermo Fisher will distribute Bio-Techne products across Europe (Photo courtesy of 123RF)")
