Current Trends in the Laboratory

iSTAT

Give an example of Lab-on-a-chip (LOC).

Analytical Stage

In most laboratory settings, this stage of testing is more automated and automated technology allows groups of tests called panels or profiles to be run on the same sample.

1. Improved efficiency 2. More physical plant requirements 3. More complex maintenance procedures

Enumerate. Automated instruments for clinical chemistry in large-volume laboratories have:

1. COVID-19 ANTIBODY-TESTING 2. MOLECULAR DIAGNOSTIC TESTING FOR SARS-COV2 3. ANTIGEN TESTING

Enumerate the 3 current methods of testing for COVID-19.

COVID-19 RT-PCR Test

a real-time reverse transcription-polymerase chain reaction test and is the gold standard for Covid-19 diagnosis

Taq polymerase

an enzyme originally isolated from a thermophilic bacteria called Thermus aquaticus.

Semi-automated urine analyzers

They can provide automated reading of urinalysis test strips in medium to high throughput settings.

True

True or False: According to WHO, studies show that the majority of patients develop antibody response only in the second week after the onset of symptoms.

True

True or False: An advantage of some point-of-care tests using whole blood is that they can be performed on blood samples obtained by finger stick rather than venipuncture.

False

True or False: Based on the reagents, IgG, IgM, and IgA can only be detected separately.

True

True or False: FDA has not yet authorized the use of neutralization tests for SARS-CoV-2.

Electronic Health Record (EHR)

A digital record of health information Streamlined sharing of updated, real-time information to move with them Access to tools that providers can use for decision making

Electronic Medical Record (EMR)

A digital version of a chart Not designed to be shared outside the individual practice The patient record does not easily travel outside the practice Mainly used by providers for diagnosis and treatment

Automated Critical Value Notification System (ACVNS)

When it identifies clinically significant changes in cardiac troponin (used in the diagnosis of myocardial infarction) results, when the software identifies the first increase of cardiac troponin, it sends an alert message to the laboratory information system. The performing lab professional then calls the patient's nurse, who informs the attending physician The software also sends the results to the physician through the EMR

yey!

(Just answer yey!) Fun Fact: The drive-thru testing being conducted in Manila and other areas of NCR like Pasig are using Enhanced Chemiluminescence Immunoassays (ECLIA) and the antibody testing available in the CDUH laboratory is also using CLIA.

yey!

(Just answer yey!) Fun fact: For antigen tests, there may be false positive results because they could occur if the antibodies on the test strip also recognize antigens of viruses other than COVID-19 such as from human coronaviruses that cause the human cold.

Denaturation

separation of two strands in the DNA double helix by raising the temperature of the mixture.

Autosomal recessive

A disease may be this which is a disease present in an individual only if two copies of the outer gene are inherited and couples who both carry this same genes have a 1 in 4 or 25% chance with each pregnancy to have a child with that disease.

CDC Influenza SARS-CoV-2 Multiplex Assay

A test that will detect two types of influenza viruses (A and B) and SARS-CoV-2 at the same time.

Multiple Methods of Analytical Detection

An analyzer with a dual function open system that can perform chemistries and enzyme-linked immunosorbent assays (ELISA) and they are also currently working towards integrating Chemiluminescence immunoassays (CLIA) and ion-selective electrode capabilities as well.

Onboard QC Management

An analyzer with an internal quality assurance system that performs more than 40 automatic checks during sample and calibration cycles to verify the correct performance of sensors, reagents, fluidics, and electronics.

Diverse assay Menu

An analyzer with more than 85 assays that cover general chemistries, esoteric or specialized tests, special proteins, therapeutic drugs and drugs of abuse

Point-of-care tests (POCT)

Are lateral flow devices that detect IgG or IgG and IgM, or total antibody in serum, plasma, or whole blood.

volume of testing performed on the instrument and their ability to connect to instrument clusters or a larger laboratory automation system

Automated systems in clinical chemistry can be differentiated based on two characteristics; What are those?

Digital microscopic analysis

Automated systems to classify and count cell types based on a digital scan of the slide or a representative number of regions of interest

Hemolysis

Avoidance of sudden accelerations and decelerations as well as the use of proper packing materials inside the carriers can help to minimize _____________.

Coulter principle

Based on changes in electrical impedance produced by non-conductive particles suspended in an electrolyte solution as they pass through a small aperture between electrodes. RBCs, white blood cells, and platelets are identified by their sizes.

Antigen Testing

Based on the detection of Viral Proteins expressed by the SARS-CoV-2 virus in respiratory tract specimens such as nasopharyngeal and nasal swabs.

Maxi Code Quick Response (QR) Code Data Matrix Aztec Code

Before, we had the traditional one-dimensional bar code. Now, the two-dimensional bar codes available include the following: (Enumerate)

Pseudovirus neutralization tests (pVNT)

Can be performed in BSL-2 laboratories depending on the VSV strain used

Time from onset of illness The concentration of the virus in the specimen Quality of the specimen and how it is processed A precise formulation of the reagents in the test kits

Enumerate Factors that affect Antigen Testing.

Binding antibody testing

Depending on the complexity of assays, these tests can be performed rapidly in a field setting or in a few hours in a laboratory.

No

Does WHO recommend the use of antigen detecting rapid diagnostic tests for patient care? Yes or No?

LIS/LIMS

Ensures not only the privacy and security of patient data, but also the quick retrieval of patient data when diagnosis and early treatment are tantamount to life-saving efforts.

Automated Testing Algorithms (ATA)

Includes evidence-based order sets for the health system's Computerized Physician Order Entry (CPOE) software

Virus neutralization tests (VNT)

Includes plaque-reduction neutralization test and microneutralization.

Neutralizing antibody detection

Involves incubating serum or plasma with live virus followed by infection and incubation of cells.

Lab-on-a-chip (LOC)

It has been successfully used in the detection of analytes such as electrolytes.

Human RNase P

It is for the detection of human nucleic acids and is used as a control for sample integrity.

Diagnostic testing

It is used to identify or confirm the diagnosis of a disease or condition in a person or a family. It gives a yes or no answer in most cases

Pre-analytical stage Analytical stage Post-analytical stage

Laboratory automated processes occur in 3 stages. What are these stages?

Elongation or Extension

New strands of DNA are made using the original strands as templates. A DNA polymerase enzyme (usually Taq polymerase) joins free DNA nucleotides together.

LIS connectivity

One of the functionalities of automated systems in clinical chemistry. Some machines have a file import/export method that makes data transfer versatile and adaptable to many of its systems.

Capsule pipelines or pneumatic tube transport

Pneumatic tubes are also known as?

Amplification

Polymerase Chain Reaction testing can detect even very small amounts of viral genetic material in a sample by duplicating it many times over through a complex laboratory process called _____________.

Virus neutralization tests (VNT)

Requires BSL-3 laboratories and may take up to 5 days to complete.

Microchip

Researchers have developed this that is embedded in the plastic of the test tube and used to store all relevant information such as when and where a sample is from, as well as the patient's name.

COVID-19 ANTIBODY-TESTING

Serologic tests detect waning or past SARS-CoV- 2 virus infection indirectly by measuring the host's humoral immune response to the virus. This is through the detection of antibodies produced by the host over days to weeks after infection with the virus.

LIS/LIMS

Serve to reduce repetitive manual lab tasks, allowing for greater hands-free data processing and results management.

Neutralizing antibody detection

Since this uses a live virus, testing will require either BSL-3 or BSL-2 laboratories, depending on what form of the SARS-CoV-2 virus is used. Is this Binding or Neutralizing antibody detection?

1. Point-of-care tests (POCT) 2. Laboratory tests

The 2 categories of Binding antibody detection tests are: (use numbering)

3 primer and probe sets - detect three regions in the SARS-CoV-2 nucleocapsid (N) gene 1 primer and probe set - detect human RNase P (RP) in a clinical sample

The RT- PCR test uses: (enumerate)

Glucometers

The most common POCT devices that have been made small and portable with improved specimen collection techniques. These are used for capillary blood sugar level.

Antigen Testing

The patient sample is added to a solution that breaks up the virus, freeing its proteins for recognition by the test

Nanotechnology

The science of building miniature devices out of very small particles such as individual atoms, molecules, viruses, or cells

Age, Nutritional status, Severity of disease, and certain medications or infections (like HIV) that suppress the immune system.

The strength of antibody response depends on:

Neutralizing antibody detection

The test determines the functional ability of antibodies to prevent infection of virus in vitro

Diagnostic testing Predictive genetic testing Pre-symptomatic genetic testing Carrier testing Prenatal diagnosis Preimplantation studies Newborn screening

There are several types of genetic testing, what are they?

Preimplantation studies

These are used following In-vitro Fertilization (IVF) to diagnose a genetic disease or condition in an embryo before it is implanted into a mother's uterus

Binding antibody detection

These tests use purified proteins of SARS-CoV-2, not a live virus, and can be performed in lower biosafety level laboratories.

Pneumatic Tube System (PTS)

They are used for transporting solid objects such as blood bags and blood samples in evacuated collection tubes

Predictive genetic testing

This determines the chances that a healthy individual with or without a family history of a certain disease might develop that disease. This is for those diseases which manifests themselves in adulthood such as some types of cancer, cardiovascular diseases and some single gene disorders

Pre-symptomatic genetic testing

This is used to determine whether persons who have a family history of disease but no current symptoms, have the gene alterations associated with the disease.

Newborn screening

This is performed in newborns in-state public health programs to detect certain genetic diseases for which early diagnosis and treatment are available.

Carrier testing

This is performed to determine whether a person carries one copy of an altered gene for a particular disease.

Prenatal diagnosis

This is used to diagnose a genetic disease or condition in a developing fetus and includes maternal serum screening, ultrasound or sonograms, amniocentesis, chorionic villus sampling, and percutaneous umbilical blood sampling

True

True or False: For the test sample, studies have shown that nasopharyngeal swabs showed a higher positive rate than oropharyngeal swabs and that is why currently existing protocols require both types of swabs to be collected to increase the sensitivity of the test.

True

True or False: Nanotechnology is already used in tests for tuberculosis and colon cancer and another potential application is in drug administration.

True

True or False: Pneumatic Tube System - Although it expedites specimen transport, close attention to the design is necessary to prevent hemolysis of the specimen.

True

True or False: The RT-PCR remains to be the gold standard for the diagnosis while the antigen and antibody testing are used for epidemiological surveillance.

False (RNA)

True or False: The SARS-CoV-2 virus is a DNA virus.

True

True or False: The antigen test results can be read by a device that measures either the fluorescence signals (FIA) or lateral flow immunochromatography type of test.

True

True or False: The clotting of blood in specimen collection tubes and their subsequent centrifugation as well as the transfer of serum to secondary tubes require a LONG time to complete.

True

True or False: The cycle is repeated many times (usually 20-30 times as most processes using PCR need large quantities of DNA) and it takes 2-3 hours to get a billion or so copies.

False

True or False: The use of whole blood in the assay system LENGTHENS the specimen preparation time.

False

True or False: These home testing products are relatively expensive, over-the-counter diagnostic and monitoring kits and devices.

False

True or False: Two-dimensional bar codes offer the prospect of encoding more data in LARGER formats and having greater reliability than one-dimensional bar codes

False

True or False: Usually, it is the IgM that rises LAST after IgG but in SARS-CoV-2 infections, the two antibody classes arise nearly simultaneously

Flow cytometry

Typically count cells often stained with supravital or fluorescent dyes that travel in suspension one by one past a laser light source. Information is derived through measurement of light scatter when a cell is struck by the laser beam (both side-scattered and forward-scattered light). Light scatter is then used to estimate cell shape, cell size, cellular granularity, nuclear lobularity, and cell surface structure.

1. Coulter principle 2. Flow cytometry 3. Digital microscopic analysis

Under Hematology, Analyzers that perform a complete blood count has been automated through the use of: (use numbering)

Pneumatic Tube System (PTS)

Under the Pre-analytical stage, it provides rapid specimen transport and are reliable when installed as point-to-point services.

Laboratory tests

Use ELISA or CLIA methods for antibody detection using blood samples.

Virus neutralization tests (VNT)

Use a SARS-CoV-2 virus from a clinical isolate or recombinant SARS-CoV-2 expressing reporter proteins.

Pseudovirus neutralization tests (pVNT)

Use recombinant pseudoviruses (like vesicular stomatitis virus, VSV) that incorporate the S protein of SARS-CoV-2

nephelometry, turbidimetry, and a wide range of immunoassays which includes ELISAs, FIAs (Fluorescence Immunoassays), and CLIAs.

What are included in the antibody-based detection methods?

Automation Information Technology Laboratory measurement or Testing technology

What are the 3 major technological innovations?

The speed of the test, which can provide results in minutes Can generally be produced at a lower cost than PCR tests

What are the advantages of Antigen Testing?

Spike glycoprotein (S) and Nucleocapsid phosphoprotein (N)

What are the antigenic targets used in Antibody testing?

The presence of antibodies cannot be equated with an individual's immunity from SARS-CoV-2 infection. Some tests may exhibit cross-reactivity with other coronaviruses and produce false positives. Some persons may not develop detectable antibodies after coronavirus infection. Based on current data, WHO does not recommend the use of antibody detecting rapid diagnostic tests for patient care but encourages the continuation of work to establish their usefulness in disease surveillance and epidemiologic research.

What are the limitations of Covid-19 Antibody testing?

Pregnancy testing kits

What are the most common examples of a home testing?

1. Denaturation 2. Annealing 3. Elongation or Extension

What are the steps in conducting an RT-PCR test? (enumerate by numbering)

1. Binding antibody detection 2. Neutralizing antibody detection

What are the types of Antibody testing? (use numbering)

1. Virus neutralization tests (VNT) 2. Pseudovirus neutralization tests (pVNT)

What are the types of Neutralization Tests?

Support the development of vaccines Determine the proportion of a population previously infected with SARS-CoV-2 and therefore may have higher rates of herd immunity Assist in determining who may qualify to donate convalescent plasma.

What are the uses of Covid-19 Antibody testing?

1. LIS/LIMS 2. Electronic Health Record (EHR) 3. Electronic Medical Record (EMR) 4. Automated Critical Value Notification System (ACVNS)

What comprises Information Technology of laboratory services? (numbering)

Monoclonal antibody

What do you call the antibody that traps the solution containing viral proteins in antigen testing which recognizes a specific segment of the SARS-CoV nucleocapsid protein?

Chemiluminescence immunoassays

What does CLIA stand for?

Clinical and Laboratory Standards Institute

What does CLSI stand for?

Enhanced Chemiluminescence Immunoassays (ECLIA)

What does ECLIA stand for?

Enzyme-linked immunosorbent assays

What does ELISA stand for?

Reverse transcriptase

What enzyme converts the RNA template of SARS-CoV-2 into complementary DNA?

However, antigen tests may be fast and cheap but they are not as sensitive as the molecular diagnostic test. This is because antigens detected are expressed only when the virus is actively replicating. Therefore, such tests are only best used to identify acute or early infection, and how well these antigen tests work also depends on several factors.

What is the disadvantage of Antigen Testing?

When there is no viral genetic material in the sample so amplification will not occur and no fluorescent markers will be detected

What's the indication of the RT-PCR test to be a positive result?

When there is viral genetic material in the sample so amplification occurs releasing enough fluorescent markers to be detected

What's the indication of the RT-PCR test to be a positive result?

July 2, 2020

When did the US FDA issue an emergency use authorization for the use of the CDC Influenza SARS-CoV-2 Multiplex Assay?

Algorithm

a process/set of rules to be followed by a computer

Genetic Testing

a type of medical test that identifies changes in chromosomes, genes, or proteins.

Pharmacogenomics

aims to develop rational means to optimize drug therapy, with respect to the patients' genotype, to ensure maximum efficiency with minimal adverse effects.

Pharmacogenomics

analyzes how the genetic makeup of an individual affects his/her response to drugs.

Pneumatic tubes

are systems that propel cylindrical containers through networks of tubes by compressed air or by partial vacuum.

Home Testing

decentralized and physicians may not receive the test results unless they are provided manually by patients or entered into shared Internet-based data-monitoring systems.

More physical plant requirements

distinct disadvantage; floor space, ventilation, water supply, electrical and drainage requirements

Gold nanoparticle bio-barcode assay probe

for the detection of prostate-specific antigen after radical prostatectomy for prostate cancer. This is especially useful since the available PSA immunoassays lack sensitivity.

Automated or semi-automated ion-selective electrodes

have been incorporated into automated systems to provide test results within minutes. This is now commonly used for assays of electrolytes and other analytes

C. difficile GDH assay

is qualitative enzyme-linked fluorescent assay technique that detects the C. difficile antigen, glutamate dehydrogenase (GDH), as a screen for the presence of C. difficile in stool specimens from persons suspected of having C. difficile infection.

Cloud LIMS

laboratory information management software built exclusively on cloud-based technology or web based

Fully automated urinalysis analyzers

perform chemical analyses, analysis of sediment or cellular elements using flow cytometry

iSTAT

rapid analyzer that rapidly analyzes very low volumes of blood and it contains arrays of electrodes deposited on silicon cartridges to form a biosensor.

IQA system

reports a test parameter only when all the 40 performance checks are successful.

Lab-on-a-chip (LOC)

the miniaturization of analytical devices that integrate several laboratory operations such as PCR and DNA sequencing into a single chip on a very small scale (only a few mm to a few cm2). It can handle extremely small fluid volumes down to picolitres.

Annealing

the temperature is lowered and the primers (small pieces of specifically engineered genetic material) are introduced and bound to the isolated viral genetic material initiating amplification. Primers mark the target sequence that will be amplified.

Improved efficiency

they can handle more tests per hour with a larger capacity for specimens and reagents