IBTS - Environmental Monitoring Flashcards
Why is EM important?
To ensure that the environment in which the products are produced has a reduced risk of contamination
What does EM do in the IBTS
Carry out tests which count the bioburden on surfaces and in the air
This allows the IBTS to spot and track trends e.g. seasonal bacteria etc
Collecting this data allows for corrective action and prevention methods e.g. retraining a staff member associated with bacterial contamination
What are the four types of trends seen
Shift (temporary) e.g. once off contamination by E.Coli
Shift permanent e.g. increased contamination associated after hiring new staff member
Cyclical e.g. seasonal contamination
Drift e.g. as months get warmer etc
What products is EM particularly vital for?
Transplantation of sterile cells and tissues e.g. Heart valves and stem cells
Information on EM is required in order for the cells/tissues to be released for transplantation
Who might look for EM info during audits and why might they require these?
The Health Products Regulatory Authority might look for EM data to ensure nothing unexpected occurred during processing, and that there is compliance with the regulations
e.g. heart valves cannot be released without certification that the cleanroom is within spec
Where is EM carried out in the IBTS
Collection clinics -> donor arms, equipment, fridges etc
Cleanrooms -> blood processing equipment
Blood storage and issue areas
What does MODA do>
Used to keep track of EM samples e.g. plan scheduling EM, acquisition of samples, incubation, result entry etc
What is the LIS management system used for EM in the lab
MODA -> Laboratory information management system
How do we monitor the blood donation clinic
Apheresis clinic monitored on a 4-week rotation where different beds/locations are monitored
Normal clinics monitored monthly and equipment is rotated etc
Venepuncturists carry out a competency check every 6 months
We will carry out a bacterial check on the donors arm post disinfection of the donor site -> assuring VP is trained correctly
How is the donors arm disinfected
We use Chloraprep or Frepp
Both contain 1.5mls of 2% chlorohexidine gluconate (IPA)
What is the acceptance criteria for EM on the donor’s arm
,5 Colony forming units on 95% of the arm
What are the main sources of contamination in cleanrooms?
Surfaces, Equipment and Water - 10%
Air -15%
Personnel - 75%
How can personnel contaminate cleanrooms?
Skin flakes, hair, cosmetics, bacteria, viruses, skin cells, cellulose fibres from clothes etc
How do we control our clean rooms
We do so by minimising the generation and retention of particles in the room using several different methods depending on the grade of sterility of the room
How do we grade cleanrooms
Grade A - cleanest to grade D being a normal laboratory room
Give some ways of maintaining a cleanroom
Hand washing & hygiene
Tacky mats - Dycem mats
Gowning and behaviour of staff
Cleaning/use of sterile materials
Design of the room/regular cleaning etc
Heating, Ventilation, Air conditioning systems (HVAC)
How does HVAC differ in a grade A/B lab than in a grade D/C
Turbulently ventilated in a grade D/C
Laminar flow/unidirectional in a grade B/A
How our the grade D labs controlled in the IBTS (not normal hospital)
Non sterile garments e.g. labcoat
Blue scrubs
Mobcap
Gloves
Lab coat
Lab safety shoes/shoe covers
How do we control a grade A/B lab
Sterile garments - spacesuit scrubs
y-irradiation sterilised:
- boots, hood, suit, long sleeved tunic, face mask and gloves
- goggles and socks (yellow) also introduced
- staff must be recertified every 6 months
How are the different labs in the IBTS graded
Production = grade D
Tissue bank;
- depending on area (grade A, B and D)
- Cryobiology: (grade A, B and D)
How do we carry out particle counts
We use a Lasair particle counter
This counts particles between 5um and 0.5um
Detects both viable and non-viable particles
Measures 1 m^3 of air
Testing is performed at several locations in a single room
What do we use settle plates for
Used to detecct viable particles -> alive
We use TSA for bacteria and SDA for yeasts and moulds
The settle plates are exposed for 4 hours
What is TSA
Tryptic Soy Agar for bacterial sedimentation EM
What is SDA
Sabouraud Dextrose Agar for yeasts and moulds sedimentation Em
How do we EM using active air
We use an SAS active air sampler to detect viable particles which works by pulling air down onto an agar plate
How would you decide what type of EM to use
Contact plates are used for flat surfaces -> they contain a grid so results can be expressed as CFU/cm^2
Swabs are used for irregulat surfaces -> TSA pour plates used for enumeration i.e. swab on plate for counting
What do the TSA contact plates contain
Naturalisers like Tween and lecithin which work against disinfectants
What kind of microbes would you expect to detect?
Gram positive microorganisms (skin commensals)
Gram negative microorganisms (eyes, ears and mucus)
Common environmental fungi anf yeasts
Give some examples of skin commensals you would expect to find
S. epidermidis
S. capitis
S. hominis
S. aureus
Micrococcus species
Bacillus species
Give some examples of GNM you would expect to find
P. aeruginosa
E. coli
Give some examples of yeasts/moulds you would expect to find
Penicilium species
Candida species
What kind of water do we use
Water sterilised with ozone (O3) formed by passing oxygen through UV light
Why do we use ozone sterilised water
Its an oxidising agent toxic to most waterborne organisms
No odour produced from it
Creates less harmful by-products than chlorination
How do we control our growth media
The European Phaemacopeia lists the tests, incubation condiitons and the microorganisms required for each type of media
We use ATCC control strains e.g. S, aureus or Candida albicans as our controls
10-100 CFU/ml inoculum made up for each media -> growth must be detectable on the media within a certain timeframe
Passing criteria (>50% and <200%)
What is ATCC
American Type Culture Collection
What product in particular required screening
Platelet screening has been a mandatory tests since 2004
How many platelets does the IBTS supply
25,000 platelets (pooled + apheresis)
What is the shelf life of platelets
7 days
Talk about storage of platelets
Stored at 22 degrees celsius
- too low promotes phagocytosis
- too high increases bacterial risk
Agitation and breathable bags necessary
What are the three main sources of platelet contamination
Bacteria present on/in the skin of the donor
Donor bacteraemia - in blood
Contamination of the blood pack itself
What percentage of platelets are contamination
Between 1 in 1000 and 1 in 3000
What safety measures are in place for platelet donations
HLQ Questionnaire
Arm cleansing
Diversion pouch on pack
Overnight hold of blood
Leucodepletion of pack
Bacterial testing
Donor notification -> if donor feels unwell 24 hours after donation
Platelets not issued until 60 hours after collection as most significant BacT positives <24 hours post collection
What does the BacT do
An automated blood culture system
Screens for bacteria in all platelets
Allows for the continuous monitoring of platelets over their shelf-life i.e. continuous monitoring
Also used for testing products associated with suspected transfusion reactions
How does the BacT system work
Two samples taken - one in an aerobic bottle and one in an anaerobic bottle
Any microorganisms multiply in the media generation CO2
As CO2 increases, the sensor in the bottle turns from grey/green to yellow
Colorimetric sensor detects this colour change by measuring reflected light every 10 minutes
REES alarm connected to BacT instrument to notify on call personnel
What bacterial has the longest BacT time?
Cutibacterium - takes 97 hours
What is done with positive blood cultures
Gram stain and subculture on to Blood (aerobic and anaerobic), chocolate and SDA agar
Retest any platelet and associated products
Isolates sent for ID in St. James’
Isolates frozen and stored indefinitely
Follow up on the donor -> treat infection
Follow up on venepuncturists -> training records etc
What is the majority of bacteria identified
Propionibacterium (45%)
Coag neg staph (33%)
How would a patient present with bacterial contamination
Febrile reaction -> increase in temp of 1.5 degrees
Chills
Hypotension (decreased bp)
All occuring within 2 hours of transfusion
How is bacterial contamination in transfusion treated
Stop transfusion
Take patient blood cultures
Administer IV antibiotics
Perform inspection and testing on blood pack
How do you examine the blood pack
Look for:
- abnormal colour
- hameolysis
- clots/leaks
Microbiology testing on:
- pack contents
- segment line
- administration line ( not done anymore)
How do you follow up on bacterial transfusion reactions
Confirm Transfusion transmitted infection -> same bacteria in blood pack and blood culutre
Molecular typing to establish strain and route of transmission
Follow up with donor depending on the type of bacteria identified
