Mohammad Rahbar (Ph.D) Professor of clinical Microbiology Department of Microbiology Iranian...

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  • Mohammad Rahbar (Ph.D) Professor of clinical Microbiology Department of Microbiology Iranian Reference Health Laboratory ,Tehran Iran

  • QC program The laboratory director is primarily responsible for QC and QA programs. However all laboratory personnel must actively participate in both program

  • Positive Patient Outcome in the Microbiology laboratoryReduced length of stayReduced cost of stayReduced turn-around time for diagnosis of infection Change to appropriate antimicrobial therapyCustomer ( physician or patients )satisfaction

  • The basic elements of QC

    Specimen collation Standard operating procedures (SOPM). Personnel Proficiency testing Antimicrobial Susceptibility tests Eminence of QC procedures Maintenance of QC stocks Patients reports

  • SPECIMEN COLLECTION AND TRANSPORT The laboratory is responsible for providing instructions for the proper collection and transport of specimens .These instructions should be available to the clinical staff for use when specimens collected.

  • Written collection instructionTest selection criteriaPatients selection criteriaTiming of specimen collection (e.g. before antimicrobial are administration)Optimal specimen collection siteApproved specimen collection methodSpecimen transport medium

  • ContinueSpecimen transport time and temperatureSpecimen holding instructions if it cannot be transported immediately (e.g. hold at 4C for 24 hours)Availability of test (on site or sent to reference laboratory )Hours test performed (daily or batch)Turn-around timeResult reporting procedures

  • Information should be filled

    Patient nameHospital or laboratory numberOrdering physicianWhether the patient receiving antimicrobial therapySuspect agent or syndrome

  • Criteria for Unacceptable SpecimensUnlabeled or mislabeled specimensUse of improper transport medium Excessive transport timeImproper temperature during transport or storageImproper collection site for test requestSpecimen leakage out of transport containerSera that are excessively hemolyzed ,lipemic, or contaminated with bacteria

  • Standard Operating Procedure Manuel (SOPM) The SOPM is considered part of QC program. The SOPM should define test performance , tolerance limits, reagent preparation, required quality control ,result reporting and references. The SOPM should be written in CLSI format and must be reviewed and signed annually by the microbiology director.

  • Continue.. The SPOM should be available in the work area .It is the definitive laboratory reference and is used often for questions relating to individual test .Any obsolete procedures should be dated when removed from SPOM and retained for at least 2 years.

  • PersonnelIt is laboratory director's responsibility to employ sufficient qualified personnel for the volume and complexity of the work performed. Document competency and training twice a yearContinuing education program should be provided All documentation should maintained in personnel file.

  • Proficiency TestingLaboratories are required to participate in an external proficiency testing (PT) The laboratory must maintain an average score of 80% to maintain licensure in any subspecialty area.The laboratory's procedures, reagents, equipments and personnel are all checked in the process of proficiency testing.

  • PT or External quality controlProvide laboratory management with an insight into their performanceImprove both local and national standardsReveals unsuspected area of difficultyProvides an educational stimulate for improvementsActs as a check on the efficacy of internal quality control proceduresDemonstrates to colleagues and customers a commitment to quality

  • Performance Checks

    InstrumentEquipment logs should contain the following informationInstrument name, serial number, and date put useProcedure and periodicity( daily, weekly, monthly) for routine function check)

  • ContinueAcceptable performance rangesInstrument function failure ,including specific details of steps taken to correct the problems (corrective action) Date and time of services requests and response.Date of routine preventive maintenance (PM) which should follow manufactures recommendations.

  • Continue..Maintenance records should be retained in the laboratory for the life of instrument. Specific guidelines regarding periodicity of testing for autoclaves, biological safety cabinets ,centrifuges ,incubators, microscope, refrigerators ,freezers, water bathes , heat blocks and other microbiology laboratory can be found in reference books .

  • Quality control of Culture Meda Commercially prepared culture mediaIn house prepared culture media (User-Prepared)

  • Commercially Prepared Culture Media The CLSI subcommittee on media quality control collected data over several` years regarding the incidence of QC failure of commonly used microbiology media Based on its finding the subcommittee published a list of media that did not require retesting in the user's laboratory if purchased from a manufactures who follow CLSI guidelines

  • Continue..The laboratory must inspect each shipment forCracked mediaExcessive bubblesClarityHemolysisFreezingUnequal fillingVisible contamination

  • User-Prepared and ,commercially prepared MediaQC forms for user-prepared media should contain:The amount of preparedThe source of each ingredientThe lot numberSterilization methods

  • Continue..The preparation date The expiration date (Usually 1 month for agar plate and 6 month for tube media)The name of prepareAll user prepared colures media also should checked for:

  • Continue.Proper colorDepthSmoothnessHemolysisExcessive bubblesContamination

  • Sterility CheckA representative sample of the lot should be test for sterility;5%of any lot is tested when a batch of 100 or fewer unit is received and maximum of 10 units are tested in large batches. Sterility is routinely checked by incubating the medium for 48 hours at the temperature at which it will be used.

  • Performance testingWhen medium dose need to quality controlled because it was prepared in house (in the laboratory) or because it is complex, several basic rules must be followed :All media must be tested before useEach medium must be tested with organisms expected to give positive reaction as well as with organism expected either not to grow or produce a negative reaction

  • Continue..The medium should be tested for sterility and pHThe organisms selected for QC should represent the most fastidious organisms for which the medium was designed Testing technique should be different for primary plating media that for biochemical or subculture media. Primary plating media should be tested with dilute suspensions of organisms, whereas biochemical media can be tested with undiluted organismsQC testing should be performed according to CLSI recommendationExpiration date must be established

  • Media failure logDate 2/14/2014Media Urea AgarLot# In house preparation 2/13/2014Expiration date 6 month from preparationQuantity 2 racksFailure failure to give proper reaction with P.mirabilis

    Action taken QC repeated with P.mirabilis failed All tubes discarded .New Urea agar preparedTechnologist MAR

  • Use of Stock CulturesTo operate a quality control program in Microbiology lab. stock culture must be maintained by all laboratories .They are available from many sources.Commercial sourcesProficiency testing sourcePatients isolatesAmerican Type Culture Collection (ATCC)

  • Continue When quality control testing appears have failed, it is usually the stock culture rather than the test itself that has failed. Organisms may mutate with repeated sub culturing. for best results ,a stock culture should be grown in a large volume of broth ,then divided among enough small freezer vials to last a year.

  • Continue..With this technique a new vial can be removed from the freezer weekly so that organism do not have to be continually subcultured .An organism may need to be subcultured twice after thawing to return it to a healthy state. Media selection for freezing is at the discretion of individual laboratories but should not contain sugar. If organism utilize sugar while being maintained ,the acid products that result may kill organism with time.

  • Popular Media for Stock CulturesSchaeler broth wit glycerolChopped meat (anaerobes)Tryptic Soy agar deeps (at room temperature) Tryptic Soy Broth (with 15 % Glycerol ) at -70 CCystein-tryptic agar (CTA) without carbohydrates

  • ContinueNonfastidious (rapidly growing), aerobic bacterial organisms can be saved up to 1 years on TSA slants .Long term storage of aerobes or anaerobes can be accomplished either by lyophilizartion (freeze drying) or freezing ,at -70C.Frozen ,no fastidious organism should be thawed ,reisolated and refrozen every 5 years; fastidious organisms should be thawed reisolated ,and refrozen every 3 years. Stock isolated may be maintained by freezing them in 10% skim milk ,Trypticase Soy Broth (TSB) with 15% glycerol .

  • Stain and Reagents Containers of stains and reagents should be labeled as to contents, concentration ,storage requirements, date prepared (or received) date placed in service ,expiration date, source (commercial manufacture or user prepared) and lot number .All stains and reagents should be stored