Download HEMO-AEROBIC Culture medium for the isolation of fastidious

HEMO-AEROBIC
Culture medium for the isolation of fastidious aerobic and
facultative anaerobic microorganisms in blood samples.
DESCRIPTION
HEMO-AEROBIC is supplied in bottle that contains a very nutritive liquid medium for growing fastidious, aerobic and facultative anaerobic microorganisms from blood.
For an optimum use the product is available in three presentations:
Adults Use: 100 mL bottle with 70 mL of culture medium.
Pediatric Use: 50 mL bottle with 30 mL of culture medium.
Neonatal Use: 20 mL bottle with 5 mL de of culture medium.
The bottles for Adults, Pediatric and Neonatal use are endowed with a grey rubber stopper perforable with the inoculation needle , an aluminium cap and a protective green stopper.
CONTENT OF THE PACKAGE
On the basis of its use, the content of each package is the following:
Adults Use:
• 6 bottles with 70 mL of Brain Heart Infusion broth enriched.
• 1 instructions sheet
Pediatric Use:
• 6 bottles with 30 mL of Brain Heart Infusion broth enriched.
• 1 instructions sheet
Neonatal Use: (also recommended for children under 2 years)
• 6 bottles with 5 mL of Brain Heart Infusion broth enriched.
•
1 instructions sheet
NOT INCLUDED PRODUCT
Sterile syringe or other device for blood collection
Disinfectant for skin
Slides, coverslips, microscope
Culture medium for subcultures
PRINCIPLE OF THE METHOD
Blood samples must be collected from the patient with rigorously aseptic technique and sterile devices. The so-obtained sample is inoculated into the hemo-culturing bottle and mixed
with the liquid medium whose formulation increases the growth of fastidious, aerobic and facultative anaerobic microorganisms from blood.
Brain Heart Infusion and peptone in the broth provide amino acids and proteins for microorganisms growth.
Sodium chloride maintains the osmotic balance of the medium.
Sodium phosphate bibasic is the buffer of the medium and dextrose is a carbon source
Sodium-polyanethol-sulphonate inhibits coagulation, neutralizes bactericide effect of human serum, prevents fagocytosis and partially inactives some antibiotics (streptomycin,
kanamycin, gentamicin and polymyxin B).
CULTURE MEDIUM FORMULA (g/L)
BRAIN HEARTH INFUSION BROTH ENRICHED
Brain Hearth Infusion…............... 17,0
Sodium Phosphate Bibasic............. 2,5
Peptone....................................... 10,0
p- Aminobenzoic Acid......................0,05
Dextrose ..................................... 2,0
Sodium-polyanethol-sulphonate......0,3
Sodium Chloride............................5,0
Final pH 7.4 ± 0.2
COLLECTION AND STORAGE OF THE SAMPLE
Whenever possible the blood sample must be collected before starting antimicrobial therapy, following a practice operation using strictly aseptic and sterile equipment.
It is very important that the blood sample is obtained with the least possible risk of accidental contamination. The area of entry of the needle must be prepared and disinfected as good
laboratory practice, through the use of alcohol or other disinfectant for the skin.
Do not touch the skin after disinfection. Perform the blood collection, taking care not to contaminate the blood and skin disinfectant. The sample volume depends on the
recommendations of experts in the field, and according to the patient's condition at the time of sampling. It indicates the volume as a minimum content of 8 - 10 mL of blood in the case
of adults, 1 - 3 mL in the case of a sample in children, and of 0.5 - 1 mL in neonatal samples (2). The use of different presentations of Hemo Aerobic can be evaluated by medical personnel,
depending on the volume of blood obtained and the age of the patient. The bottles of Hemo Aerobic may be used for the isolation of infectious agents from other types of samples, in
particular from sterile liquids.
TEST PROCEDURE
a) Inoculation technique
1.Observe the broth in the bottle and check there are not signs of contamination and deterioration.
2.Lift the protective plastic cap and disinfect the external side of the rubber stopper.
3.Perforate the rubber stopper with the syringe needle and aseptically inject the blood: negative pressure in the bottle will allow the intake until 10 mL of blood in the bottle for Adult use,
until 3 mL in pediatric bottle and 1 mL in Neonatal bottle, included children under 2 years, (NEONATAL presentation).
4.Carefully mix blood with culture medium in the bottle.
5.Write patient's name and other informative data on the label and incubate at 36 ±1 °C during 16-24 hours.
b) Subculturing Technique
1.Shake the bottle gently to homogenize the mixture of broth inside.
2.Draw with a sterile syringe a small amount of broth mixture which must be inoculated into the culture media designed for the isolation of potentially pathogenic bacteria
3.Put a drop of the mixture of broth from the syringe and deposit it in a glass slide for microscopic observation using Acridine Orange or Gram stain.
4.Examine at the microscope (100X) to observe the presence and the morphology of microorganisms.
5.Perform observation at microscope and subcultures at 16 hours and after 24, 48 and 72 hours. If there is no growth of microorganisms, prolong the incubation time of the bottle at 36 ±
1 °C until 7 days, before performing final subculture and considering negative the result.
Rev.1/03.01.2012
HEMO-AEROBIC
Culture medium for the isolation of fastidious aerobic and
facultative anaerobic microorganisms in blood samples.
QUALITY CONTROL
Each batch of HEMO- AEROBIC is subjected to the quality control using the following reference microorganisms:
Escherichia coli
ATCC 25922
Pseudomonas aeruginosa
ATCC 27853
Staphylococcus aureus
ATCC 25923
Streptococcus agalactiae
ATCC 13813
Klebsiella pneumoniae
ATCC 13883
Staphylococcus epidermidis
ATCC 12228
Candida albicans
ATCC 10231
Enterobacter aerogenes
ATCC 13048
Streptococcus pneumoniae
ATCC 6305
Haemophilus influenzae
ATCC 10211
Haemophilus parainfluenzae
ATCC 7901
Enterococcus faecalis
ATCC 19433
Listeria monocytogenes
ATCC 35152
Streptococcus pyogenes
ATCC 19615
FACTORS THAT MAY INVALIDATE THE RESULTS
Bacterial growth in the bottles can be due to the skin flora or to contamination during the collection or inoculation. In these cases a second blood culture or clinical importance of the
isolated microorganism will indicate clinically significant infections. Take in consideration that it can be collected 2-3 quotes of blood sample in different points to increase the detection of
bacteremia.
LIMITS AND WARNINGS
HEMO-AEROBIC can promote the growth of a wide variety of microorganisms.
Nevertheless in clinical use some microorganisms can fail to growth for the following reasons:
- The strain is particularly fastidious.
- The microorganism was not present in the blood sample.
- Presence in the blood of an inhibitory concentration of any antimicrobial.
- Some microorganisms do not grow in the recommended incubation time. In some clinical cases may require an incubation period longer than 7 days.
CAUTIONS
The product, HEMO-AEROBIC, cannot be classified as hazardous under current legislation, nor does it contain harmful substances in concentrations ≥1%; it therefore does not require a
Safety Data Sheet to be available.
HEMO-AEROBIC must be used in the laboratory by properly trained personnel, using approved aseptic and safety methods for handling pathogenic agents.
STORAGE
Store at 10-25 °C in the original packaging. Keep away from sources of heat and avoid excessive changes in temperature.
In such conditions, the product will remain valid until the expiry date indicated on the label. Do not use beyond that date. Waste if there are signs of deterioration.
DISPOSAL OF USED MATERIALS
After use, HEMO-AEROBIC and material that has come into contact with the sample must be decontaminated and disposed of in accordance with the techniques used in the laboratory
for decontamination and disposal of potentially infected material.
REFERENCES
1.
Finegold S M and Martin W J. (1982) Diagnostic Microbiology 6th ed. Published C.V. Mosby Co. St. Louis p.42.
2.
Hinder S M, Sawhney D and Swaine D. 2nd European Congress of Clinical Microbiology 1985. Abstract 12/2.
PRESENTATION
Product
Code
Package
HEMO-AEROBIC
MH0197
6 bottles X 70 mL
HEMO-AEROBIC PEDIATRIC
MH0198
6 bottles X 30 mL
HEMO-AEROBIC NEONATAL
MH0199
6 bottles X 5 mL
TABLE OF SYMBOL
Batch number
Do not reuse
Manufacturer
Do not use if package is
damage
Temperature limitation
Catalogue Number
Fragile, handle with care
Used By:
Caution, consult
accompanying documents
in vitro diagnostic
Medical Device
C.P.M. sas – Via degli Olmetti, 5 – 00060 (Zona Ind.le) Formello, (ROMA), Italia - Tel.
+39/06.90.400.126 - +39/06.90.75.531 - Fax. +39/06.90.405.270 – www.cpmsas.it – [email protected]
Rev.1/03.01.2012