Oxford Jr Trainee BMS Flashcards
How do your skills meet the job description of this role?
My skills align closely with the requirements of the trainee BMS position, ensuring I can effectively meet the demands of the role
- attention to detail; for instance I consistently ensure that patient samples accurately match with the request cards minimising errors and ensuring precise results. My meticulous nature guarantees the integrity of all data and samples I come across
- I am diligent in making sure the reagents and test kits, for instance the legionella urine test kits, that we use to process patient samples have been acceptance tested, and also verifying the expiry dates of reagents and media to ensure they’re suitable for use, thereby maintaining high standards of accuracy and reliability
- my experience and knowledge of the analysers for instance kiestra and test procedures within our laboratory reduces the learning curve and enhances productivity especially when the opportunity arises for me to learn more about the other analysers
- I am well versed in the techniques and methodologies necessary for processing patient samples, including the use of automation, which ensures timely and accurate results
-I possess the initiative to tackle tasks independently while also being a strong team player contributing to a collaborative and efficient work environment
- I am adept at following standard operating procedures and complying with ISO 15189:2012 standards, ensuring that all processes meet regulatory and quality requirements
- I am committed to ongoing professional development and I am currently undertaking the IBMS pre-registration portfolio, Demonstrating my dedication to advancing my skills and knowledge
-my excellent communication skills enable me to convey information clearly and effectively, while maintaining strict confidentiality and adhering to data protection protocols
-I embody the NHS trust values, as well as the OUH trust values in my work, ensuring compassionate and respectful care while striving for excellence in all tasks
-I handle criticism constructively, seek feedback for continuous improvement, learn quickly and perform well under pressure. My flexibility allows me to adapt to changing circumstances and demands.
With my skills and attributes, I am confident in my ability to contribute effectively to our department and excel in this trainee role
OUH trust & values
OUH Nhs trust is one of the largest in the country that provides a range of general and specialised clinical services
OUH values:
-excellence
-compassion
-respect
-delivery
-learning
-improvement
Its aim is to provide excellent care, with compassion and respect, for example by putting patients at the heart of what is done and going the extra mile & following through with commitments
It also aims to deliver, learn & continuously improve for example, delivering the best clincial and teaching research whilst also monitoring & assessing performance by learning from successes and setbacks
Why is confidentiality important?
Confidentiality in the NHS is important as all staff have a legal duty to respect it and understand the confidentiality procedures applicable to the trust that they are working in
Confidential information relating to service users must never be shared unless it’s needed for the safe and effective care of the individual. It is important to obtain consent before sharing any confidential information, and if you’re ever unsure, seek advice
Final year project
It was based on the overuse of hand sanitisers and the development of Antimicrobial resistance
The aim of the project was investigate whthefe there was a variation in the Antimicrobial activity of commercial alcohol free and alcohol based hand sanitisers against bacteria commonly found on the hands (Enterococcus faecalis, pseudomonas aeruginosa, e. Coli, s. Aureus and s. Epidermidis) and that are known to cause illnesses, and to determine if resistance of chosen bacteria to hand sanitisers can arise
This was determined via a range of assays:
- growth of overnight cultures—> use of sterile technique to introduce innoculum of pure culture to sterile nutrient broth
- agar well diffusion —> determine efficacy of hand sanitisers by measuring zones of inhibition
- MIC —> determine MIC
- sub inhibitory concentration exposure assay——>evaluate if bacterial species had the ability to acquire antimicrobial resistance to hand sanitisers at low concentration. Afterwards, the MIC was tested again to see if there was any changes/resistance
The final result showed that, although alcohol-based hand sanitisers showed a decreased antimicrobial efficacy against organisms, it was the only hand sanitiser to show a decreased generation of resistance. Whilst alcohol free hand sanitisers suggested that their overuse could lead to the development of potential resistance.
General data protection regulation/GDPR
The way in peoples personal information is used is covered by a law known as GDPR
Healthcare professionals who provide care, maintain records about our health and treatment or care that we have received previously. These records help to provide us with the best healthcare and treatment. These records can be electronic paper based or a mixture of both
The information is stored and can only be used for management audit purposes. However, it is only available to and used by those involved in our care. We have the right to know what information that is held about us and if you would like to see what information is held, we could always ask. patients have the right to prevent confidential information from being shared or used for any purpose other than providing care , except personal circumstances.
As a staff member, I would ensure I follow the code of practice & rules applicable to the laboratory team and I will make sure I would never leave confidential information or records lying around. Never discuss anything about a patient unless it is within the permitted realms of my work with authorised personnel. Also within the boundaries of the code of conduct
What is the difference between hazard and risk?
Hazard is something that can cause harm. for example, chemicals, walking up a ladder or electricity/ electrical devices
Risk is the chance either high or low, that any hazard can actually cause somebody harm 
What is SOP
Standard operating procedures
Step-by-step guide on how to perform a task. All staff required to perform a particular task must do so in the same way as everybody else by following the guidelines outlined.
SOP benefit patients the most, as they can be confident that their sample will be handled in the same way, and undergo the same procedure as everyone else’s
Why are SOP important?
They are good for quality management inconsistency
Ensure all experiments being performed are done in a consistent manner to produce reproducible results.
Also important from a safety aspect as written instructions would include information about potential hazards and how these can be mitigated
Sterilisation vs disinfection
Sterilisation is a thorough, sterilisation or removal of all microbes present
Disinfection is the reduction in the total number of microbes below risk level
What is COSHH?
Control of substances hazardous to health regulations
It is the law that requires employers to control substances that are hazardous to health and perform risk assessments
All packages and samples must clearly labelled as hazardous
Tell me why you think you would be a good fit for this role
I believe I would be an excellent fit for this role within our department for several reasons
- My undergraduate degree in biomedical science has provided me with a solid foundation in various scientific disciplines, including microbiology, I have a deep understanding of microbial physiology, genetics and the role of microorganisms in health and disease
- My laboratory experience throughout university, and my hands-on experience of working within this laboratory, have eqipped me with practical skills that are directly applicable to this role. During this time I have become proficient in essential laboratory techniques, sample data entry, processing, culturing, use of various laboratory instruments and laboratory waste management, all whilst adhering to safety protocols
-I am well versed in aseptic techniques, handling and processing clinical specimens and the operation of microbiology specific equipment. My familiarity with standard operating procedures and quality control measures ensure that I can contribute to maintaining high standards of accuracy and reliability in laboratory results
- my academic and practical experiences have honed my analytical and problem-solving skills. I am adept at troubleshooting experimental issues and interpreting data
-I understand importance of teamwork in a laboratory environment. My experience has taught me to communicate effectively with colleagues share responsibilities and collaborate to achieve common goals. I am also comfortable taking initiative and working independently when needed.
-I have a genuine interest in microbiology and it’s applications in healthcare. Notably, my final year project at university was in microbiology. I am eager to expand my knowledge and stay updated with the latest advancements in the field. My passion drives me to contribute positively to the department and continuously improve my skills.
-precision and accuracy are critical in biomedical science, and I pride myself on my meticulous attention to detail whether it’s recording data, preparing samples or performing experiments. I ensure that every task is carried out with the utmost care.
Given my educational background experience, technical skills and passion for microbiology, I am confident that I would be a valuable asset to our team and excel in this trainee role
Sell yourself to us in one minute
With my strong educational background in biomedical science, and my hands-on experience within this laboratory, I bring a solid understanding of microbiology and a proven ability to perform essential laboratory techniques with precision.
My practical skills, combined with a keen attention to detail and a passion for the field make me a quick learner and relevant and reliable team member. I am adept at troubleshooting, interpreting data, and maintaining high standards of quality.
My enthusiasm for microbiology and commitment to continuous learning ensure that I will contribute positively to the department and excel as a trainee BMS
IQA
Internal quality assurance
IQA involves the internal processes and systems put in place to ensure the accuracy, reliability and consistency of test results.
This includes:
standard operating procedures for all testing processes
regular staff training and competency assessments
proficiency testing and inter- laboratory comparisons
routine review and updating of protocols to align with current standards and scientific advancements
internal audits to assess adherence to SOPs and identify areas for improvement
EQA
External quality assurance
EQA involves assessments conducted by external bodies to validate the quality and reliability of the laboratories testing. This can include:
Participation in external proficiency testing programs where the laboratory results are compared with those from other laboratories
accreditation with recognised bodies for example, ISO15189
external audits conducted by regulatory authorities or accreditation organisations to ensure compliance with international standards
IQC
Internal quality control
IQC involves routine checks and procedures to monitor the performance of tests and equipment within a laboratory. This can include:
Regular calibration and maintenance of laboratory equipment
Use of control samples, positive and negative controls, in each batch of tests to ensure accuracy
Monitoring of media and reagents for contamination and performance
Documentation and analysis of control data to identify trends in deviations
NEQAS
United Kingdom national external quality assessment service
NEQAS provides external quality assessment schemes for laboratories in the United Kingdom and globally. It aims to improve the quality of laboratory procedures and insures the reliability of test results. Key aspects include
regular distribution of reference samples to participating laboratories for testing.
Comparison of laboratory results with the expected results, and those from other laboratories
Feedback on performance, highlighting areas of excellence and those needing improvement
Educational resources in guidance to help laboratories improve their practices
Horizontal audits
A horizontal audit focuses on specific aspects or processes across different departments or sections. This type of audit may often follow an examination audit where are finding has suggested that more thorough auditing is required. This type of audit is typically used for auditing the quality management system, transport, pre-/post examination processes, and IT
for example:
Reviewing the sample collection and handling procedures across all departments to ensure uniformity
Checking compliance with biosafety protocols in various sections of the laboratory
Vertical audit
Vertical audit examines all aspects of a single department or process from start to finish. It covers pre-examination, examination and post examination processes. It’s aim is to check that the SOP accurately reflects what is being done on the bench to ensure that it’s correctly written and current and to check the person carrying out the task has a good understanding of the processes involved thereby reflecting the departments, training and assessment of competence.
for example:
Auditing the entire workflow of the bacteriology section from sample receipt to result reporting to ensure compliance with standard
Detailed examination of the mycology sections procedures for culturing and identifying fungi
Purpose of quality systems
Quality insurance and control measures such as audits, IQC, IQA, EQA, and NEQAS help laboratories maintain high standards of accuracy and reliability and safety in their testing procedures
Departments quality system
The department utilises several internal procedures to check that our procedures remain fit for purpose. These include: data analysis
acceptance testing
internal quality control
internal quality assurance
event reports
non-compliance
internal audits
quality improvement
quality objectives
Acceptance testing
Acceptance testing is conducted to determine if equipment, reagents, and consumables tested against specific materials achieve the expected outcome/s. Within the department, this can include:
-growth or no-growth
-reactivity or non-reactivity when tested against NCTC/ACTC strains (collection of bacterial strains to provide microbiologists with reliable source of authenticated strains
-Reactivity or non-reactivity when tested against commercial IQ material
GLP
Good laboratory practice it is based on basic principles, which should be second nature to all laboratory staff.
-Lab coats to be worn at all times within the laboratory and to be placed individually on hooks in the corridor when not in use
-Gloves and eye protection to be worn when appropriate. when moving into another part of the laboratory, remove one glove to ensure that you do not contaminate doorhandles. never answer or use the phone whilst wearing gloves
-Ensure all cuts and abrasions are covered with waterproof plasters
-Wearing sensible shoes —> flat and cover the entire floor
-Spillages to be cleared up appropriately by the person responsible for the spillage, and not to be left for the next person to use the bench or piece of equipment
-Do not leave equipment or benches dirty
- tie long hair back, so it does not swing forward
- all sharps or objects that has the potential to puncture an autoclave bag to be disposed of in a sharps container
-No eating, smoking or running in the laboratory
-Never put your fingers, pens or pencils into your mouth eyes
-Washing your hands before you leave the laboratory, and after suspected contamination
Categorisation of pathogens
The organisms we work with in laboratory are classified into different levels to help us handle them in the correct environment.
They are categorised into 4 groups
Group 1 - unlikely to cause human disease (Lactobacillus)
Group 2 - can cause human disease, may be hazardous to employees, but it is unlikely to spread to the community and there is usually effective prophylaxis for treatment available (S. aureus, Candida, enterobacteriaceae, neiserria gonorrhoea
Group 3 - can cause severe human disease and maybe a serious hazard to employees. It may spread to the community but there is usually effective prophylaxis for treatment available (mycobacterium tuberculosis, salmonella typi, shigella, HIV, bacillus anthracis, sporadic creutzfeldt Jakob disease
Group 4 - causes severe human disease, and as a series has a temporary is it is likely to spread to the community and there’s usually no effective prophylaxis for treatment available (Ebola virus, variola virus, haemorrhagic fever viruses, Lassa fever
Containment level 3
Containment level 3 suite is where category 3 pathogens that can cause sever human diseases are handled.
This suite has engineering controls such as:
Specialised air handling system that ensures a directional airflow. Air flows from clean areas to potentially contaminated areas with air being filtered through a high efficiency particulate air (HEPA) filters before being exhausted.
The lab is designed to be airtight with sealed windows, floors, walls, and ceilings to prevent any leakage of airborne contaminants
Entry strictly controlled and requires multiple levels of security, often, including anterooms and airlocks to minimise the risk of contamination when entering or leaving the lab
Lab staff wear powered air purifying respirators and other forms of respiratory protection, as well as enhanced PPE, including full bodysuits, gloves, and face protection
All work with infectious material is conducted within a class 1 biosafety cabinet to prevent exposure and contamination
There are rigorous protocols for decontaminating work surfaces, equipment and waste
Chloros
Used for cleaning analysers such as BD VIPER & BD KIESTRA and wiping the outside of COVID swabs
Not recommended to disinfect tuberculosis materials
Precept granules
Used for larger spills and is poured on & mixed into spill, taking care to avoid chlorine fumes given off. Ensure to increase ventilation by opening windows and avoid inhalation of fumes.
Tristel
Tristel is active against bacteria, including mycobacterium species, fungi, spores and viruses. It is less corrosive than chloros and is used in the disinfection of the category 3 laboratory cabinets and incubators.
Gram staining
Gram staining is a fundamental technique used in microbiology to classify bacteria into two major groups, gram-positive and gram-negative. This method was developed by Hans Christian gram in 1884 and is based on the differences in the cell wall structures of bacteria.
It is useful to know the size and arrangement of bacterial cells in stained film as organisms are traditionally classified according to the Gram staining reaction
Differences in gram-positive and gram-negative bacteria cell wall composition account for gram staining differences. Gram-positive cell walls contain thick layers of peptidoglycan with numerous teichoic acid cross-linking, which resist decolourisation.
In aqueous solutions crystal violet dissociates into positively and negatively charged ions (CV+ & CL-) that PENETRATE THROUGH THE CELL WALLS OF GRAM-POSITIVE AND GRAM-NEGATIVE ORGANISMS. POSITIVELY CHARGED IONS/CV+ interact with negatively charged components of bacterial cells staining them purple.
When iodine is added, which is negatively charge, it interacts with a positively charged crystal violet ions to form large crystal violet – iodine complexes within the cytoplasm and outer layers of cell
Decolorizing agents, such as ethanol or acetone, interact with lipids, or both gram-positive and gram-negative membranes
The outer layer gram negative bacteria is lost, leaving thin peptidoglycan layer is exposed. The addition of ethanol or acetone causes the cell walls to become leaky and allows crystal violet-iodine complexes to be washed from the cells. When adding counter stains, such as neutral red, that is positively charged, it allows the gram-negative bacteria to stain red/pink.
However, highly cross-linked and multilayered peptidoglycan walls of gram-positive bacteria dehydrates after adding ethanol or acetone therefore, the ethanol/acetone traps large crystal violet and iodine complexes within the cell, allowing the cell to remain purple
Gram staining method
1) a smear is prepared on a slide either from patient samples or directly from cultures by using an inoculation loop to transfer a drop of suspended culture onto the slide. Using the inoculation loop, the culture is spread into an even film and then is left to air dry
2) the culture is then fix onto the microscope slide using a hot plate for 10 minutes
3) The slide is then flooded with crystal violet which is left on for about 30 seconds.
4) excess stain is gently rinsed off with running tap water. The goal is to wash of the stain without losing the fixed culture.
5) the slide is then flooded with grams iodine. This method is known as fixing the dye, which is left for about 30 seconds and then rinsed with running tap water.
6) a few drops of the decoloriser is added to the slide. The decolouriser used is grams differentiator. This step is known as solvent treatment. Decolourisation only takes 2 to 3 seconds and you should be careful not to over decolourise. Therefore you should stop adding decolouriser as soon as the solvent is not coloured as it flows over the slide.
7) the slide is then immediately washed with water
8) smear is counterstained with neutral redfor about one minute. After that it is then washed off and bloated dry.
9) the slide is then examined using oil immersion objective at x50 or 100 to observe cell morphology and gram reaction
PCR
Polymerase chain reaction
PCR is a widely used technique in microbiology to amplify and detect specific DNA sequences. It allows for the rapid and precise identification of micro organisms, including bacteria, viruses, fungi, and parasites.
PCR involves the amplification of a specific DNA segment through a series of temperature regulated steps
It is made up of the following components :
Template DNA - the DNA extracted from the microbial sample that contains the target sequence to be amplified
Primers -short synthetic DNA sequences that are complementary to the regions flanking the target DNA sequence. They provide starting points for DNA synthesis
DNA polymerase - an enzyme that synthesise’s new DNA strands by adding nucleotides to the primers. Taq polymerase is commonly used as it remains active at high temperatures.
Nucleotides (dNTPS) - the building blocks (adenine, thymine, cytosine & guanine) used by the DNA polymerase to construct new DNA strands
Buffer solution - maintains the optimal pH & ionic conditions for the activity of DNA polymerase
PCR steps
Denaturation - the reaction mixture is heated to around 94 to 98°C to separate the double strand of DNA into single strands
Annealing - the temperature is low at 2:50 to 65°C to lie primers to bind/anneal to the complimentary sequences on the single-stranded DNA
Extension - the temperature is raised to 72°C. The optimal temperature for taq polymerase, which synthesises new DNA strands by adding nucleotides to the annealed primers.
The steps are repeated for 20 to 40 cycles, leading to an exponential increase in the number of copies of the target DNA sequence
Advantages & limitations of PCR
Advantages
-sensitivity, PCR can the text very low amounts of DNA, making it possible to identify pathogen is present in small quantities
-specificity, are use a specific primers allows, for the selective amplification of target sequences reducing the likelihood of false positives
-speed, PCR can produce results within a few hours which is much faster than traditional culture, beast methods
-Versatility, PCR can be adapted to detect a wide range of micro organisms, including those that are difficult of slow to culture
Limitations:
-contamination risk, the high sensitivity of PCR makes it susceptible to contamination, leading to false positive results
-Complexity, require specialised equipment and trained personnel to perform and interpret results
-Quantification challenges, standard PCR is primarily qualitative, providing presence or absence information without indicating the amount of pathogen present
Uncertainty of measurement
Pipetting - calibration, maintenance, competence
Centrifugation- calibration, maintenance
Media & reagents- acceptance testing, expiry dates, IQC & EQA
Incubation time & temperatures - measuring, calibration
Samples- minimal turnaround time from collection to processing, storage, specimen containers
Personnel- training & competency
Name cat 2 organisms
Campylobacter species
Clostridium difficile
Non pathogenic strains of e. Coli
Haemphilus influenzae
Candida albicans
Cryptosporidium species
Rubella virus
Name cat 3 organisms
Bacillus anthrasis
Brucella abortus
Mycobacterium leprae
Yersinis pestis
Plasmodium falciparum
dengue virus
Clinical importance of gram staining
Aids in prompt and appropriate therapeutic decisions
Differentiation of bacteria aids in the identification of type of bacteria causing infection
Results give empirical antimicrobial therapy. For example gram + cocci might suggest streptococcal or staphylococcal infections while gram - might suggest infections caused by e.coli or pseudomonas species
Knowing gram reaction helps choose appropriate antibiotics as gram + and gram - have different susceptibilities
