Download Applied Biosystems 7500 Maintenance and Calibration

Applied Biosystems 7500
Maintenance and Calibration
When to Calibrate the ABI 7500
When you install the system. You must perform all
calibrations in the following order:
ROI,
Background,
Optical, Dye,
instrument verification.
Every 6 months, or as often as necessary, depending on
instrument use.
After replacing the lamp.
IMPORTANT! After every ROI calibration, you must
perform a background calibration, optical calibration, dye
calibration, and instrument verification.
Maintenance Menu
Region of Interest Calibration
The ROI calibration maps the positions of
the wells on the sample block so that the
software can associate increases in
fluorescence during a run with specific
wells of the plate. Because the instrument
uses a set of optic filters to distinguish the
fluorescence emissions gathered during
runs, you must generate a calibration
image for each individual filter to account
for minor differences in the optical path.
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Troubleshooting
Condition: The ROI image is faint or you
cannot generate a successful calibration.
The sample block may be in its lowered
position.
If the CCD Control Settings in the ROI Inspector
displays Block Up, click Block Up to raise the
sample block.
Check that the heated cover assembly is pulled
all the way forward to ensure that the tray can be
pushed in properly
Background Calibration
Background Calibration
If the instrument did not complete the background calibration
run:
Repeat the calibration.
If the problem persists, contact Applied Biosystems.
If the background calibration failed:
One or more wells of the background plate produced spectra that
exceed the maximum limit for background spectra on the Applied
Biosystems 7500/7500 Fast Real-Time PCR System.
Repeat the calibration using the same background plate.
If the calibration fails again, repeat the calibration using a different
background plate.
If the calibration fails again, determine the source of the
contamination.
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Optical Calibration
Optical Calibration
As per ROI calibration.
Same plates and handling conditions.
Compensates for the extra deep red filter
found in 7500 series.
Spectral Calibration
Spectral Calibration
Spectral Calibration
Spectral Calibration
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Lamp Status
Lamp status
The Lamp Status/Replacement dialog box displays:
Condition – Indicates one of the following conditions:
Good – The lamp is functioning well and does not need to be
replaced. Click Close.
Failed – The lamp bulb must be replaced. Click Close, then
replace the lamp.
Change Soon – The lamp usage is above 2000 hrs; Applied
Biosystems recommends that you replace it. Click Close, then
decide whether or not to replace the lamp.
Usage (Hours) – The total number of hours that the lamp has
been illuminated.
Lamp Current – The output current of the lamp in amperes (A).
Low current can indicate a potential future failure of the lamp.
Date Last Replaced – The date of the last lamp replacement.
When to Calibrate the ABI 7500
When you install the system. You must perform all
calibrations in the following order:
ROI,
Background,
Optical, Dye,
instrument verification.
Every 6 months, or as often as necessary, depending on
instrument use.
After replacing the lamp.
IMPORTANT! After every ROI calibration, you must
perform a background calibration, optical calibration, dye
calibration, and instrument verification.
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Victorian Infectious Diseases
Reference Laboratory
Quality Assurance and
Good Laboratory Practices
Research Institute of Tropical Medicine
Manila, 4 December 2012
Bruce Thorley
• Major public health facility
- AS ISO 15189:2009
• Infectious disease
- laboratory diagnosis
- outbreak investigation
- surveillance
• State Programs
• National Programs
• International Programs
Victorian Infectious Diseases Reference Laboratory (VIDRL),
Australia
Why is GLP Necessary?
We want to have confidence that
laboratory results we receive are timely,
accurate, sensitive, appropriate
Diagnostic and therapeutic laboratories
work within a highly regulated
framework
Quality Control and
Quality Assurance
QC: internal monitoring, parallel testing,
maintain competency
QA: external monitoring, participation in
appropriate proficiency program
Good Laboratory Practice
Generates high quality, reliable test
results
Promotes the quality & validity of test
data
Underpinned by a Quality System
concerned with an organisation’s
management
What is a Quality System?
A structured and documented management
system that describes the policies,
objectives, principles, organizational
authority, responsibilities, accountability,
and implementation plan of an organization
for ensuring quality in its work processes,
products (items), and services
U.S. Environmental Protection Agency
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What is a Quality System?
Elements of a Quality System
Quality System Manual
A structured and documented management
system that describes the policies,
objectives, principles, organizational
authority, responsibilities, accountability,
and implementation plan of an organization
for ensuring quality in its work processes,
products (items), and services
U.S. Environmental Protection Agency
Document Controlled
Laboratory Manual
Standard Operating procedure (SOP)
Section performing the test
states what the organisation does
and how it is put into practice
Defined organisational structure
Good management practices
Good documentation
Protocol for test procedures:
Detailed
Accurate
Up-to-date (document control)
Meets regulatory requirements
Document control details
Standard Operating procedure (SOP)
Standard Operating Procedure
• Clear, unambiguous, can be understood
by staff with and without experience
• Describe specific biosafety hazards and
precautions that need to be taken
• Material Safety Data Sheets (MSDS)
available for hazardous chemicals used
• Worksheet to record data
• List references associated with the
procedure
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Standard Operating Procedure
Staff training
Ensure adequate number of trained staff
are available for all procedures at all
times
Annual review of training record
(list of laboratory procedures)
“trained, in-training, not applicable”
Staff Competency
Staff competency
Staff training
Retaining expertise in test performance,
understanding the theory & practice
How do we maintain competency with
poliovirus real time RT-PCR as more
countries move to IPV?
Biosafety
AS/NZS 2243.3:2010
Safety in Laboratories
Staff immunisations:
comply with Australian Immunisation
Handbook – 9th ed. 2008
Comply with WHO Polio Reference
Laboratory
Induction of staff
Adequate training & supervision
Biorisk = biosafety + biosecurity
Part 3:Microbiological aspects and
containment facilities
Degree of hazard from microorganisms
Physical containment levels
Laboratory spills
Special equipment
Work areas, cleaning & waste
Transport of biological material
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Documentation Trail
Where was the work
performed?
Laboratory records clearly show:
why, how & by whom the test was done
who supervised
what equipment was used
results obtained
if problems were encountered & how
they were addressed
Where are the specimens
stored?
Biological Safety Cabinet Use
Laboratory Infrastructure
Laboratory Infrastructure
Messages
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TRACEABILITY
Establish a trail of data from receipt of a
specimen in the lab to reporting of the
final result
Is it documented?
Equipment Log & Maintenance
Asset register
Maintenance schedule
Cleaning roster
Calibration (eg micropipettes every 3 months)
Special instructions
Electrical safety check
Equipment
Equipment Messages
Enrolment in Annual
Proficiency Panels
International Accreditation
Accreditation requirement
Education
Verify methods & procedures
Identify problems
Assess performance
Comparison with peers
Demonstrate competence
International Organization for
Standardization (ISO)
National Association of Testing Authorities
(NATA)
Established in 1947, first laboratory
accreditation body
NATA determines a facility’s technical
competence based on the relevant
international standard
Source: Proficiency Testing Australia
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Standard for Human Pathology
Testing
Accreditation Site Review
ISO 15189
International quality management system standard
AS ISO 15189:2009 (ISO/IEC 17025)
Medical laboratories - Particular
requirements for quality and
competence.
WHO Polio Reference Laboratory
Accreditation Checklist
An accreditation is a fact finding
mission not a fault finding mission
Introductory meeting
Evaluation of the technical
competence & examination of the
quality management system
Exit meeting
Quality System
Equipment maintenance
SOPs
Lab manual
Quality system manual
Document control
GLP
Proficiency panels
Staff training
Result recording & reporting
Internal audits
Defined management structure
Quality control
Biosafety
Method validation
Physical environment
Maintaining a Quality System
Improvements to the system
Improvements to the system
Maintaining a Quality System
Time
Time
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Improvements to the system
Maintaining a Quality System
Temperature chart 2006
Next accreditation visit
Time
Temperature chart 2010
Temperature chart 2012
Temperature Monitoring
PCR Reagent Storage
• Temperature probes in air/glycerol
• On-call scientist notified of alarm out of hours
• Remote log-in to assess situation
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Store Enzymes in a Benchtop
Cooler
Label Reagent Storage Boxes
Use a Black Box to Store
Real Time PCR Probes
VIDRL Quality Systems Group
International Transport of
Infectious Substances
Internal Audits
Internal (peer) review of a specific item across the
institute
- Are staff training records / equipment calibrations / laboratory manuals up to date?
- How does each lab record batch numbers and expiry dates of
reagents?
Investigate whether section management /
procedures conform to the Quality System Manual
Quality System Notification
Corrective Action Request
Key process for system maintenance & improvement
Can assist with establishing institutional policy
Need to be qualified & re-certified every 2 years
Carriage of infectious substances on board an
aircraft in
- checked-in luggage
- hand luggage
- on a person
is strictly prohibited by law
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Infectious Substance
Category A
Infectious Substance
Category A: Indicative list UN2814
An infectious substance which is transported
in a form that, when exposure to it occurs, is
capable of causing permanent disability, lifethreatening or fatal disease in otherwise
healthy humans or animals.
WHO: Guidance on Regulations for the Transport of
Infectious Substances 2011-12
Infectious Substance Category B
Proper shipping name:
UN3373 Biological substance, Category B
Shipment Packaging
Contents list
Secondary receptacle
Dry ice label
An infectious substance which does
not meet the criteria for inclusion in
category A
Examples: stool specimens for poliovirus culture
Customs declaration, import
permit & dangerous goods
declaration
Australian Quarantine and
Inspection Service (AQIS)
UN3373 or
UN2814 label
UN packaging
specification
Australian Quarantine and
Inspection Service (AQIS)
Minimise the risk of exotic pests & diseases entering
Australia
Check with AQIS whether an item requires an import
permit
Restricted material must be handled at a Quarantine
Approved Premise (QAP) by trained persons
QAP must comply with AS/NZS 2982.1:1997
(Laboratory Design and Construction) and AS/NZS
2243.3:2002 (Safety in Laboratories).
Spot-checks & issue Corrective Action Request
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Quality System
Equipment maintenance
SOPs
Lab manual
Quality system manual
Document control
GLP
Proficiency panels
Staff training
Result recording & reporting
Internal audits
Defined management structure
Quality control
Biosafety
Method validation
Physical environment
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