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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

As per ROI calibration.

Same plates and handling conditions.

Compensates for the extra deep red filter found in 7500 series.

Optical Calibration

Spectral CalibrationSpectral 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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Quality Assurance and Good Laboratory Practices

Research Institute of Tropical Medicine

Manila, 4 December 2012

Bruce Thorley

Victorian Infectious Diseases Reference Laboratory (VIDRL),

Australia

Victorian Infectious Diseases

Reference Laboratory

• Major public health facility- AS ISO 15189:2009

• Infectious disease- laboratory diagnosis - outbreak investigation- surveillance

• State Programs

• National Programs

• International Programs

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

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

Quality Control and Quality Assurance

� QC: internal monitoring, parallel testing, maintain competency

� QA: external monitoring, participation in appropriate proficiency program

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 servicesU.S. Environmental Protection Agency

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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 servicesU.S. Environmental Protection Agency

Elements of a Quality System

� Quality System Manualstates what the organisation does

and how it is put into practice

� Defined organisational structure

� Good management practices

� Good documentation

Document Controlled Laboratory Manual

Section performing the test

Document control details

Standard Operating procedure (SOP)

Protocol for test procedures:

� Detailed

� Accurate

� Up-to-date (document control)

� Meets regulatory requirements

Standard Operating procedure (SOP)

• 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

Standard Operating 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

� 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?

Staff training

Biosafety

� 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

AS/NZS 2243.3:2010 Safety in Laboratories

� 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

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 was the work performed?

Where are the specimens stored? Biological Safety Cabinet Use

Laboratory InfrastructureLaboratory Infrastructure Messages

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� Establish a trail of data from receipt of a specimen in the lab to reporting of the final result

� Is it documented?

TRACEABILITY 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

� Accreditation requirement

� Education

� Verify methods & procedures

� Identify problems

� Assess performance

� Comparison with peers

� Demonstrate competence

Source: Proficiency Testing Australia

International Accreditation

� 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

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Standard for Human Pathology Testing

� ISO 15189International quality management system standard

� AS ISO 15189:2009 (ISO/IEC 17025) Medical laboratories - Particular requirements for quality and competence.

Accreditation Site Review

� 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

WHO Polio Reference Laboratory Accreditation Checklist Quality System

Lab manual

Biosafety

SOPs

Staff training

Quality system manual

Defined management structure

GLP

Document control

Internal audits

Proficiency panels

Method validationQuality control

Equipment maintenance

Result recording & reporting

Physical environment

Maintaining a Quality System

Improvements to the system

Time

Maintaining a Quality System

Improvements to the system

Time

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Maintaining a Quality System

Improvements to the system

Time

Next accreditation visit

Temperature chart 2006

Temperature chart 2010 Temperature chart 2012

Temperature Monitoring

• Temperature probes in air/glycerol• On-call scientist notified of alarm out of hours

• Remote log-in to assess situation

PCR Reagent Storage

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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

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

International Transport of Infectious Substances

� 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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An infectious substance which is transported

in a form that, when exposure to it occurs, is capable of causing permanent disability, life-

threatening or fatal disease in otherwise

healthy humans or animals.

WHO: Guidance on Regulations for the Transport of

Infectious Substances 2011-12

Infectious Substance Category A

Infectious Substance

Category A: Indicative list UN2814

Proper shipping name:

UN3373 Biological substance, Category B

An infectious substance which does

not meet the criteria for inclusion incategory A

Examples: stool specimens for poliovirus culture

Infectious Substance Category BShipment Packaging

Secondary receptacleContents list

Customs declaration, import

permit & dangerous goods declaration

UN packaging specification

Dry ice label

UN3373 or

UN2814 label

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

Australian Quarantine and Inspection Service (AQIS)

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Quality System

Lab manual

Biosafety

SOPs

Staff training

Quality system manual

Defined management structure

GLP

Document control

Internal audits

Proficiency panels

Method validationQuality control

Equipment maintenance

Result recording & reporting

Physical environment