AUTOMATED BLOOD CULTURE
Contents
1. Page 9 of 21 A GENERAL SAFETY CONSIDERATIONS Pathogenic microorganisms including Hepatitis B Virus and Human Immunodeficiency Virus may be present in specimens Universal Precautions and institutional guidelines should be followed in handling all items contaminated with blood or other body fluids 1 Wear gloves while handling inoculated vials 2 Perform all blood culture processing in a biological safety cabinet 3 Properly dispose of all contaminated materials Place syringes needles and other sharp contaminated materials in a puncture proof container WARNING Never attempt to recap a needle B PROCESSING NEW BLOOD CULTURES 1 ENTERING DATA AND LOADING INSTRUMENT 1 1 BACTEC 9240 9120 a Entering Data into the Computer e From the Main menu press F3 Culture Complete the following fields Patient s ID Patient s Name Accession Number Collection Date Collection Time Hospital Service KANNAN e Advance the cursor to the sequence number field With the computer s barcode scanner scan the vial barcode label The sequence number media type station number and the status field are then filled in automatically Check the protocol field Default protocol is 5 days for blood cultures and 42 days for MYCO F LYTIC NOTE To change the length of the testing protocol use the J to advance the cursor to the protocol field and enter the desired value 1 to 42 days ENTERING DATA AND2. Polyanetholesulfonate SPS Cationic and Non ionic Adsorbing Resins Carbon dioxide COz Oxygen O2 Sensor for the detection of fluorescence b Store at 2 to 25 C 9000 bc2 RevisionB 7 25 01 Page 1 of 21 9000 bc2 RevisionB 7 25 01 2 BACTEC PLUS Anaerobic F Culture Vial 3 Optimum blood volume for each vial is 8 to 10 mL 3 to 10 mL of blood is acceptable a Each vial contains 25 mL pre reduced enriched Soybean Casein Digest broth 0 05 SPS Resins CO and Nitrogen gas No Sensor for the detection of fluorescence b Store at 2 to 25 C BACTEC PEDS PLUSIF Culture Vial Optimum blood volume for each vial is 1 to 3 mL 0 5 to 5 mL of blood is acceptable a Each vial contains 40 mL Enriched Soybean Casein Digest broth 0 02 SPS Resins COs O2 Sensor for the detection of fluorescence b Store at 2 to 25 C Page 2 of 21 MEDIA CONT 4 BACTEC MYCO F LYTIC A non selective culture medium for the recovery of yeast and fungi from blood and body fluids and mycobacteria from blood specimens The optimum specimen volume is 3 to 5 mL 1 mL to 5 mL is acceptable a Each vial contains 40 mL Processed Water Middlebrook 7H9 Broth Base Brain Heart Infusion Casein Hydrolysate Supplement H Inositol Glycerol SPS Tween 80 Pyridoxal HCl Ferric Ammonium Citrate Potassium Phosphate Saponin Antifoam CO and Oo Sensor for the detection of fluorescence B M
3. other information as per facility 2 Each request slip should also have all the information above 9000 bc2 RevisionB 7 25 01 Page 6 of 21 D NUMBER AND TIMING Most cases of bacteremia are detected using two to three sets of separately collected blood cultures More than three sets of blood cultures yield little additional information Conversely a single blood culture may miss intermittently occurring bacteremia and make it difficult to interpret the clinical significance of certain isolated organisms IV QUALITY CONTROL A MEDIA Each case of media has a Quality Control certificate indicating the organisms tested and the acceptibility of those tests The laboratory may wish to test each shipment of media for performance through the use of a positive and negative vial test The positive vial should be inoculated with 1 0 mL of a 0 5 McFarland Standard of either Escherichia coli or Staphylococcus aureus prepared from a fresh 18 24 h culture This vial and an uninoculated vial should be logged into the instrument and tested The inoculated vial should be detected as positive by the instrument within 72 hours The negative control vials should remain negative throughout the entire testing protocol This verifies that the media were not subject to adverse storage or shipping conditions prior to receipt in the laboratory If either of these vials do not give the expected results do not use the media until you have contacted Technical S
4. 1 BACTEC PLUS Aerobic F and PLUS Anaerobic F Culture Vials Insert Rev PP 088E BD Biosciences 2 BACTEC PEDS PLUS F Culture Vials Insert Rev PP 091 1 BD Biosciences 3 BACTEC MYCO F LYTIC Culture Vials Insert Rev PP 162C BD Biosciences 4 BACTEC Fluorescent Series Users Manual Document Number MA 0074 BD Biosciences 5 BACTEC 9050 System Users Manual Document Number MA 0103 BD Biosciences 6 BACTEC Blood Culture Procedural Trays Document Number L 001810 A BD Biosciences 7 Isenberg H D editor in chief 1992 Clinical Microbiology Procedures Handbook American Society for Microbiology Washington DC 8 Balows A et al Manual of Clinical Microbiology 6th ed American Society for Microbiology Washington DC 1995 9000 be2 RevisionB 7 25 01 Page 20 of 21 9 Howden R J J Clin Path 1976 29 50 53 10 Recommendations for preventing transmission of Human Immunodeficiency Virus and Hepatitis B Virus to patients during exposure prone invasive procedures MMWR 1991 Vol 40 No RR 8 11 Bloodborne Pathogens Code of Federal Regulations Title 29 Part 1910 1030 Federal Register 1991 56 64175 64182 12 Reller L B P R Murray and J D McLowry 1982 Cumitech 1A Blood Culture Ill Coordinating ed J A Washington II American Society for Microbiology Washington DC BACTEC BBL FOS LUER LOK SAFETY LOK PEDS PLUS F PERSIST and VACUTAINER are trademarks of Becton Dickinson and Company
5. ATCC is a trademark of the American Type Culture Collection TECHNICAL SUPPORT For information or assistance call toll free 1 800 638 8663 selection 2 Approved By Date Effective Supervisor Date Director Date 9000 bc2 RevisionB 7 25 01 Page 21 of 21
6. ATERIALS REQUIRED BUT NOT PROVIDED VACUTAINER SAFETY LOK Blood Collection Set OR 20 mL LUER LOK syringe with a 21 gauge needle 3 mL LUER LOK syringe with a 23 gauge needle PERSIST Povidone lodine Prep 70 isopropyl alcohol alcohol pads Biological Safety Cabinet autoclave venting units mycobactericidal disinfectant microscope materials for staining slides and subculturing supplies C INSTRUMENT 1 BACTEC 9240 9120 9050 BACTEC Fluorescent Series Microorganisms if present in the blood samples metabolize nutrients in the BACTEC culture vial and release CO into the medium or utilize the oxygen in the medium The instrument monitors the fluorescence of the vial sensor which increases as CO is produced or oxygen is utilized Analysis of the rate and amount of COs produced or Op utilized enables the instrument to determine if the vial is positive i e the presumptive presence of viable organisms 9000 bc2 RevisionB 7 25 01 Page 3 of 21 2 3 Computer and peripherals The system computer stores all the system software including the application software which controls instrument operations and the user interface which enables the user to enter patient information view results print reports identify errors etc NOTE The microprocessor associated with each rack is responsible for the actual testing of the vials and positivity analysis Barcode Scanner The barcode scanner is located on the insi
7. AUTOMATED BLOOD CULTURE BACTEC 9240 9120 9050 l PRINCIPLE The detection of microorganisms in a patient s blood has diagnostic and prognostic importance Blood cultures are essential in the diagnosis and treatment of the etiologic agents of sepsis Bacterial sepsis constitutes one of the most serious infectious diseases and therefore the expeditious detection and identification of bloodborne bacterial pathogens is an important function of the diagnostic microbiology laboratory The BACTEC 9000 series of blood culture instruments are designed for the rapid detection of microorganisms in clinical specimens The sample to be tested is inoculated into the vial which is entered into the BACTEC instrument for incubation and periodic reading Each vial contains a sensor which responds to the concentration of COs produced by the metabolism of microorganisms or the consumption of oxygen needed for the growth of microorgnisms The sensor is monitored by the instrument every ten minutes for an increase in its fluorescence which is proportional to the increasing amount of CO or the decreasing amount of Oz present in the vial A positive reading indicates the presumptive presence of viable microorganisms in the vial I MATERIAL A MEDIA 1 BACTEC PLUS Aerobic F Culture Vial Optimum blood volume for each vial is 8 to 10 mL 3 to 10 mL of blood is acceptable a Each vial contains 25 mL Enriched Soybean Casein Digest broth TSB 0 05 Sodium
8. LOADING INSTRUMENT CONT e Press F10 to save the entry Enter a different Accession number for each culture 9000 bc2 RevisionB 7 25 01 Page 10 of 21 b Loading the Instrument Take the new vials to the instrument and open the instrument doors e Scan the vial s barcode label Listen for the beep indicating a successful scan and look for the station with the illuminated GREEN and RED LED Insert the vial into that station Repeat the above step for each of the new vials NOTE If you haven t already entered the patient s data into the computer be sure to write down the station number on the patient s requisition slip to use for logging patient s information into the computer at a later time Close the instruments doors NOTE Avoid placing vials into the instrument without scanning the barcode If vials are not scanned into the instrument they will become ANONYMOUS VIALS An anonymous vial must be identified as soon as possible so that the instrument can display the vial s current status i e ongoing positive etc Identifying an anonymous vial involves the removal and re insertion of vials which can result in an increase of false positive rates Refer to User s Manual Section 4 9 for identifying anonymous vials c Replacing Vial Barcode Labels NOTE Extra vial sequence barcode labels are included in the instrument start up kit These labels can be used to replace damaged or unreadabl
9. a positive or a final negative e To return a smear negative positive vial to the instrument open the doors and scan the vial s barcode using the instrument barcode scanner The original station will be indicated by FLASHING GREEN and RED LEDs changing to solid GREEN and RED LEDs NOTE Original station location will be held for three hours after a positive vial has been removed from the instrument The data for the returned positive vial are maintained until explicitly removed as a positive or a final negative 2 2 BACTEC 9050 a The system notifies the user of the presence of presumptive positive vials in several ways An audible alarm sounds if the instrument is configured to a volume of gt 0 The NEW POSITIVE INDICATOR on the front of the instruments flashes RED Page 14 of 21 9000 bc2 RevisionB 7 25 01 e The REMOVE POSITIVES soft key appears in the LCD display when the instrument door is opened e Onthe Main Status display the station with the positive vial is displayed as a filled circle with a plus sign in it and the positive total in the Summary region reflects the number of positives in the instrument To Remove the Positive Vials Press the SILENCE ALARM key to acknowledge the alarm Available only if instrument has been configured to a volume of gt 0 Open the door of the instrument and press the REMOVE POSITIVES soft key e LCD display indicates the positive vial
10. ature controller Verify that the temperature of each rack is 35 C 1 5 C and the cabinet temperature is 30 C 1 0 C Also verify that the temperature of the calibrated internal temperature probe is 35 C 1 5 C If any rack or cabinet is not within the specified temperature range call BD Biosciences Field Service at 1 800 544 7434 Check rack indicator operation by opening the instrument door and using the barcode scanner a Scan the selection ILLUMINATE GREEN RACK INDICATORS Listen for a beep indicating the scan is successful The GREEN lamps at each station should illuminate If any lamp does not the station should be removed from service Refer to User s Manual Section 4 3 for how to remove a station from service b Scan the selection ILLUMINATE RED RACK INDICATORS Listen for a beep indicating the scan is successful The RED lamps at each station should illuminate If any lamp does not the station should be removed from service Refer to User s Manual Section 4 3 for how to remove a station from service c Scan the selection ILLUMINATE FRONT PANEL INDI CATORS All four of the indicator lamps on the front of the instrument should illuminate together then one at a time If any lamp does not refer to Users Manual Section 6 7 for instructions on replacing the burned out lamp d Scan the selection AUDIBLE ALARM TEST The instrument s audible alarm will sound three times The pattern BD is also display
11. by position and the barcode number of the vial Remove the vial from the indicated station and scan the barcode label on the vial e Repeat the appropriate steps until all of the positives are removed Three audible beeps indicate all positive vials have been removed Press the EXIT soft key Close the door of the instrument Page 15 of 21 9000 bc2 RevisionB 7 25 01 NOTE f the positive vial was originally entered manually by selecting media type the LCD displays the station location only Remove the vial from the specified station and press the FORCE STATION AVAILABLE soft key NOTE A subculture and a Gram stain should be performed from each presumptive positive vial c To Return Smear Negative Positive Vials to the BACTEC 9050 e Open the door of the instrument and press the VIAL ENTRY soft key Scan the vial barcode and place the vial in the indicated station shown on the LCD display This station may differ from the original station NOTE If the vial was originally entered by scanning the barcode label the system reserves the station for re entery of the pulled positive vial for up to 3 hours The re entry feature resets positivity routines retains the previous test readings and continues to test the vial as an ongoing culture If the vial was originally entered manually the system treats the re entered vial as a new vial 3 NEGATIVE CULTURES a Negative cultures may exist as ongoin
12. de of the instrument to provide the ability to activate barcode commands and to scan vial labels for specimen identification When the door is opened and an activity is initiated the scanner is activated and is ready to read the vial barcode The barcode scanner for the BACTEC 9050 is located on the front of the instrument and is activated when the door is opened and the vial entry or vial removal soft keys are pushed lll SPECIMEN A COLLECTION 9000 bc2 RevisionB 7 25 01 1 3 SITE SELECTION a Select a different body site for each culture drawn b Avoid drawing blood through indwelling intravascular catheters unless blood can not be obtained by venipuncture Blood collected from intravascular catheters should be done with the knowledge that contamination may be an issue SITE PREPARATION PERSIST Povidone lodine Prep a Open the PERSIST package by tearing completely through at the side notches and twisting b Leave the package over the end of the swabstick to prevent gloves from becoming covered with solution c Apply PERSIST by beginning at the intended venipuncture site working in a circular motion with friction covering an area of 2 3 inches in diameter Do not return to the center of the site once swab has moved outward to the periphery Persist should be applied with friction and the site prepped 30 seconds to 1 minute d Allow PERSIST solution to air dry e DO NOT touch or palpate the area aft
13. e labels on culture vials The barcode labels contain sequence numbers which uniquely identify each vial e Ensure that the area where the defective label is located is clean and dry If the old label is wrinkled or creased peel off as much as possible to make a smooth surface on which to apply the new label e Peel off the new barcode label Verify that the new label is printed clearly and that no smears smudges or other markings obstruct the lines of the barcode e Align the new label with the old label and press the new label into place being careful not to create any bubbles or ridges in the barcode area NOTE If you replace a barcode label the system requires you to scan a media barcode located on the media barcode placard immediately after scanning the replacement vial sequence barcode It is very important that you scan the correct barcode 9000 bc2 RevisionB 7 25 01 Page 11 of 21 9000 bc2 RevisionB 7 25 01 for the type of media since the system applies different positivity criteria to the different types of media 1 2 BACTEC 9050 Press the Home Rotor soft key for easier loading This will position the rotor at the HOME position with the temperature standards in the 12 00 position a Entering New Vials NOTE NOTE Take the new vials to the instrument and open the door of the instrument Press the VIAL ENTRY soft key Scan the vial barcode by placing the vial in the alignment block in fron
14. e placing the vial into the system False positivity may occur when the white blood cell count is high It is recommended that related vials remain out of the instrument for no more than 10 minutes to minimize the possibility of the vial becoming a false positive vial BACTEC MYCOFF LYTIC vials are not selective and will support the growth of other aerobic organisms besides mycobacteria yeast and fungi Positive vials may contain one or more species of mycobacteria and or other non mycobacterial species If present fast growing organisms may mask the detection of slower growing mycobacteria yeast and fungi Subculture and additional procedures are required The consistency of microscopic morphology in BACTEC MYCO F LYTIC has not been established Inoculation of blood volumes of 1 to 5 mL are acceptable but optimum recovery is obtained with 3 to 5 mL During internal studies with less than 3 mL of blood M intracellulare M malmoense M haemophilum and M xenopi exhibited detection delays and or compromised recovery with BACTEC MYCO F LYTIC False positivity most likely will increase when the blood volume is above 5 mL Mycobacteria may vary in acid fastness depending on strain age of culture and other variables 9000 bc2 RevisionB 7 25 01 Page 19 of 21 Blood may contain antimicrobials or other inhibitors which may slow or prevent the growth of microorganisms BACTEC MYCOY F LYTIC vials are incubated at 35 C potentially prec
15. ed in the station LED s light emitting diodes followed by the instrument number BACTEC 9240 only If the alarm doesn t sound contact BD Biosciences Field Service at 1 800 544 7434 Page 8 of 21 INSTRUMENT MAINTENANCE CONT 1 3 Perform system backup a Use a 3 inch high density formatted and not write protected diskette From the main menu press F5 or Utilities menu From the Utilities menu press F5 or Backup Insert the diskette in the disk drive Press F10 to begin the backup process When the backup is completed remove the diskette and store it ina safe place Use a different diskette every day Maintain a set of seven diskettes with each diskette labeled with a different day of the week 1 4 Weekly check the air filter at the rear of the instrument Clean and replace the filter monthly Refer to the User s Manual Section 6 6 2 0 BACTEC 9050 The following checks should be performed daily 2 1 Check the temperature readout on the LCD display on the instrument Verify that the temperature of the instrument and internal temperature vial read 35 C 1 5 C 2 2 f a printer is attached to the instrument check the paper supply 2 3 Change or clean the air filters on both sides of the instrument monthly Check the filters more frequently if the laboratory s environment is dusty Refer to the User s Manual Section 6 2 V PROCEDURE 9000 bc2 RevisionB 7 25 01
16. elines recommended by your institution g The inoculated BACTEC vials should be transported as quickly as possible to the laboratory Page 5 of 21 B VOLUME The volume of blood cultured is critical because the number of organisms per mL of blood in most cases of bacteremia is low especially if the patient is on antimicrobial therapy In infants and children the number of organisms per mL of blood during bacteremia is higher than adults so less blood is required for culture 1 Children 1 to 5 mL of blood per venipuncture Transfer the entire amount to a BACTEC PEDS PLUSIF vial 2 Adult 16 to 20 mL of blood per venipuncture If it is impossible to draw the required amount aliquot as follows Amount per Amount in BACTEC Amount in BACTEC Venipuncture Plus Aerobic Vial Plus Anaerobic Vial 16 20 mL Split equally between aerobic and anaerobic vials 13 16 mL 8 mL 5 8mL 10 12 mL 5 7 mL 5 mL 5 9 mL entire blood amount 0 NOTE Optimum recovery of isolates will be achieved by adding 8 to 10 mL of blood BACTEC PEDS PLUS F 1 3 mL BACTEC MYCO F LYTIC 3 to 5 mL The use of lower or higher volumes may adversely affect recovery and or detection times C SPECIMEN LABELING 1 Each vial should be labeled with the appropriate patient information e Patient s name e Hospital number Patient ID e Patient s location room and bed e Date and time of collection e Collector s initials e Site of venipuncture e Or
17. er cleansing DISINFECTING BLOOD CULTURE VIALS Page 4 of 21 9000 bc2 RevisionB 7 25 01 C Remove the flip off caps from BACTEC culture vials Wipe top of each vial with a separate 70 isopropyl alcohol pad and allow to dry Do not use iodine to disinfect tops of vials 4 VENIPUNCTURE a Avoid touching the venipuncture site If it is necessary to touch the site f after it has been cleaned wipe your fingers with povidone iodine before touching the site When using the Blood Collection Set butterfly the phlebotomist MUST carefully monitor the volume collected by using the 5 mL graduation marks on the vial label If the volume is not monitored the stated maximum amount collected may be exceeded This condition may adversely create a false positive result due to high blood background If using a needle and syringe typically a 20 mL syringe is used for adults Draw 16 to 20 mL of blood for one blood culture set aerobic and anaerobic Aseptically inject 8 to 10 mL of specimen into each vial Aseptically inject 3 to 5 mL into the MYCO F LYTIC vial For pediatric patients a 3 mL syringe is frequently used Draw 1 to 3 mL of blood and transfer the entire amount into BACTEC PEDS PLUS F vial After all specimens have been collected from the individual use a sterile alcohol pad to remove the povidone iodine solution from the venipuncture site Continue to care for the venipuncture site following guid
18. ervices BD Biosciences at 1 800 638 8663 prompt 2 U S only It is recommended that each new shipment or lot of BACTEC MYCO F LYTIC media be tested with the ATCC control organisms identified in the chart below as positive control and an uninoculated vial as a negative control days Mycobacterium intracellulare ATCC 13950 8 to 16 Candida glabrata ATCC 15545 Cryptococcus neoformans ATCC 13690 The positive control vials should be inoculated using a 1 100 dilution of a McFarland 1 suspension of microorganisms grown on solid medium Inoculate the vial with 0 1 mL of the diluted culture The positive control vials and an uninoculated control vial should be scanned into the instrument and tested The inoculated vial should be detected as positive by the instrument within the test protocol i e range of time to detection as stated above The negative control should remain negative If expected results for Quality Control are not obtained do not use the medium and contact BD Biosciences Technical Services in the US only 1 800 638 8663 or your local BD Biosciences Representative for further assistance 9000 bc2 RevisionB 7 25 01 Page 7 of 21 B INSTRUMENT MAINTENANCE 9000 bc2 RevisionB 7 25 01 1 0 BACTEC 9240 9120 The following procedures should be performed at the start of each day s testing and recorded on the maintenance log 1 1 1 2 Check temperature readout of each rack and cabinet on the instrument s temper
19. g negatives in protocol and out of protocol negatives The BACTEC 9240 9120 instrument systems will indicate ongoing negatives by displaying the station number as solid green digits on the computer s instrument status screen Ongoing negatives are displayed as a solid circle on the Main Status Display of the BACTEC 9050 Out of protocol negatives will be displayed in flashing green digits on the BACTEC 9240 9120 computer s instrument status display Also the negative count in the summary window of the computer will flash green Out of protocol negatives in the BACTEC 9050 are displayed as a solid circle with a minus sign in the Main Status Display The summary region reflects the number of negatives in the instrument Also the REMOVE NEGATIVES soft key appears when the door is opened Page 16 of 21 d To Remove Out of Protocol Negatives from the BACTEC 9240 9120 instrument systems Open the instrument doors Scan the menu option REMOVE NEGATIVES using the instrument barcode scanner Listen for a beep from the scanner indicating the item was scanned successfully Find the station s with FLASHING GREEN LEDs and remove the vial Scan the vial barcode label using the instrument barcode scanner Listen for the beep indicating a successful scan and the GREEN LEDs will extinguish Repeat the above two steps to remove additional negatives e To Remove Out of Protocol Negatives from the BACTEC 9050 Open the door to the ins
20. he Users Manual Section 4 2 for instructions d Replacing Vial Barcode Labels NOTE Extra vial sequence barcode labels are included in the instrument start up kit These labels can be used to replace damaged or unreadable labels on culture vials The barcode labels contain sequence numbers which uniquely identify each vial e Ensure that the area where the defective label is located is clean and dry If the old label is wrinkled or creased peel off as much as possible to make a smooth surface on which to apply the new label e Peel off the new barcode label Verify that the new label is printed clearly and that no smears smudges or other markings obstruct the lines of the barcode e Align the new label with the old label and press the new label into place being careful not to create any bubbles or ridges in the barcode area e The vial can now be entered into the instrument through vial entry 2 POSITIVE CULTURES 2 1 BACTEC 9240 9120 The system will notify the presence of presumptive positive vials in several ways e An audible alarm sounds if configured and the computer Title and Cabinet windows flash The Positive Indicator Lamp on the front of the instrument iluminates yellow POSITIVE CULTURES CONT e On the computer s instrument status display the station number and the total positives in the summary window are displayed in FLASHING GREEN FLASHING RED a To Remove the Positive Vials e Pres
21. ials A contaminated sample will give a positive reading but this does not indicate a clinically significant result Such a determination must be made by the user based on such factors as type of organisms recovered occurrence of the same organism in multiple cultures patient history etc Recovery of SPS Sensitive and Fastidious Organisms from Blood Samples Because blood can neutralize the toxicity of SPS toward organisms sensitive to SPS such as some Neisseria species the presence of optimum volumes of blood based on media type benefits the recovery of these organisms To enhance the growth of SPS sensitive organisms when less than optimum volumes of blood are inoculated additional whole human blood may be added Some fastidious organisms such as certain Haemophilus species require growth factors such as NAD or factor V which are provided by the blood specimen If the blood specimen volume is 3 0 mL or less for BACTEC PLUS Aerobic F and Anaerobic F or 0 5 mL or less for BACTEC PEDS PLUS F an appropriate supplement may be required for recovery of these organisms BACTEC BRAND FOS Fastidious Organism Supplement catalog 442153 or whole human blood may be used as nutritional supplements 9000 bc2 RevisionB 7 25 01 Page 18 of 21 Non viable Organisms A Gram stained smear from a culture medium may contain small numbers of non viable organisms derived from medium constituents staining reagents immersion oil gla
22. luding the recovery of mycobacteria requiring other incubation temperatures such as M marinum M ulcerans or M haemophilum Recovery of such organisms requires additional culture methods Penicillium purpurescens and Blastomyces dermatitidis were not detectable in the BACTEC MYCO F LYTIC culture medium Hansenula anomala Exophila Jeamselmei Actinomyces bovis Rhodotorula rubra and Mucor ramosissimus exhibited inconsistent results at low inoculum levels lt 10 CFU vial with seeded culture studies Recovery of such organisms may require additional culture methods The following isolates were detected as positive is the BACTEC MYCO F LYTIC medium during internal seeded studies and or clinical trials Mycobacterium terrae Mycobacterium abscessus Cryptococcus neoformans Mycobacterium tuberculosis Mycobacterium xenopi Histoplasma capsulatum Mycobacterium avium Mycobacterium malmoense Aspergillus flavus Mycobacterium kansasii Mycobacterium haemophilum Aspergillus fumigatus Mycobacterium fortuitum Candida albicans Nocardia asteroides Mycobacterium intracellulare Candida glabrata Malasezzia furfur Mycobacterium gordonae Candida parapsilosis Trichophyton rubrum Mycobacterium szulgai Candida krusei Mycobacterium simiae Candida tropicalis Mycobacterim celatum Ajellomyces dermatitidis Any vial assigned to a new station i e in the event of a bad station should be subcultured immediately prior to placing in the new station Vil REFERENCES
23. s F2 to acknowledge the alarm Open the instrument doors and using the instrument barcode scanner scan the menu option REMOVE POSITIVES Listen for a beep indicating that the item was scanned successfully 9000 bc2 RevisionB 7 25 01 Page 13 of 21 9000 bc2 RevisionB 7 25 01 Find the station with the FLASHING GREEN and RED LEDs Remove the vial and scan its barcode using the instrument barcode scanner Listen for the beep and the LEDs will extinguish If the instrument is configured to show related vials the vial with the same patient ID and Accession number will be indicated by a solid GREEN LED If you wish to remove this vial for subculture scan its label with the instrument barcode scanner after you remove the vial from its station NOTE Related vials should be replaced within 10 minutes Repeat the appropriate steps above for each additional positive The acknowledged alarm condition is not clear until all the positive vials are removed NOTE A Gram stain and a subculture should be performed from each presumptive positive vial b To Return Smear Negative Positive Vials and Related Vials To return the removed related vial to the instrument open the doors and scan the vials barcode using the instrument barcode scanner The original station will be indicated by solid GREEN and RED LEDs Return the vial to this station The data from the related vial are maintained until explicitly removed as
24. ss slides and specimens used for inoculation In addition the patient specimen may contain organisms that will not grow in the culture medium or on media used for subculture Such specimens should be subcultured to special media as appropriate Antimicrobial Activity Neutralization of the antimicrobial activity by resins varies depending on dosage level and timing of specimen collection Studies have demonstrated that the resins present in this medium do not adequately neutralize imipenem cilastatin antimicrobial preparations Recovery of Streptococcus pneumoniae In aerobic media S pneumoniae will typically be visually and instrument positive but in some cases no organisms will be seen on Gram stain or recovered on routine subculture If an anaerobic vial was also inoculated the organism can usually be recovered by performing an aerobic subculture of the anaerobic vial since this organism has been reported to grow well under anaerobic conditions General Considerations Optimum recovery of isolates will be achieved by adding the appropriate volume of blood for the type of vial inoculated Use of lower or higher volumes may adversely affect recovery and or detection times Blood may contain antimicrobials or other inhibitors which may slow or prevent the growth of microorganisms False negative readings may result when certain organisms do not produce enough COs to be detected by the system or if significant growth has occurred befor
25. t of the scanner with the barcode facing the scanner If necessary rotate the vial slightly so the scanner will read the label The system beeps once to indicate a successful scan If the vial barcode is covered or damaged press the NO BARCODE soft key Select the vial media by using the T or J keys Press the OK soft key to confirm the media type Insert the vial into the position indicated on the LCD display If the default protocol is acceptable press the OK soft key To modify the protocol length for a particular vial select the CHANGE PROTOCOL soft key Use the 1 or J keys to select the desired protocol length Verify that all of the information displayed is correct and that the vial has been inserted into the indicated station Press the OK soft key to confirm Repeat the above step for each new vial Press the EXIT soft key when all of the new vials have been entered BACTEC 9050 VIAL ENTRY CONT NOTE Close the door of the instrument Avoid placing vials into the instrument without scanning the barcode If vials are placed into the instrument without scanning they will become ANONYMOUS VIALS An anonymous vial must be identified as soon as possible so that the instrument can display the vial s current status i e Ongoing Positive etc Identifying an anonymous vial involves the removal and re Page 12 of 21 insertion of vials which can result in an increase of false positive rates Refer to t
26. trument Press the REMOVE NEGATIVES soft key The LCD display indicates the position and the barcode number of the negative vials Remove the vial from the station and scan the barcode label NOTE If the vial was originally entered by selecting the media type the LCD displays the station location only Remove the vial from the specified station and press the FORCE STATION AVAILABLE soft key Repeat the appropriate steps until all negatives are removed Three audible beeps sound when the removal process is completed e Press the EXIT soft key e Close the door of the instrument 9000 bc2 RevisionB 7 25 01 Page 17 of 21 4 PROCESSING AN INSTRUMENT POSITIVE VIAL a Remove the vial from the instrument and place in a biological safety cabinet b Invert the vial to mix the contents c Observe Universal Safety Precautions to vent each presumptive positive blood culture vial Use a venting needle BBL Venting Units catalog 271056 d Remove aliquot from the vial for stain preparations Gram and or AFB e Subculture vials according to the Gram stain and or AFB stain results f Report preliminary results only after stain preparation g Perform identification and susceptibility of organism s grown on solid media according to your laboratory protocol VI LIMITATIONS Contamination Care must be taken to prevent contamination of the sample during collection and inoculation into the BACTEC v
Download Pdf Manuals
Related Search