COLLECTION - Tube no.
2 is for micro, gram stain and culture, if with 4th tube use
- properly collected specimen is absolutely crucial to quality the 4 th tube for better exclusion of skin contaminants.
diagnostic information and patient care. - Never refrigerate
- Centrifuged- use the sediments for smears and culture
SAFETY - Common pathogens found in csf samples: H. influenzae, N.
- Universal precautions are followed throughout the collection and meningitides
handling process.
- Persons collecting or handling specimens should wear gloves and THROAT AND NASOPHARYNGEAL SWABS
a laboratory coat. - Most abundant normal flora- Viridans streptococcus
- Eye protection should also be worn if splashing is a potential - Most common pathogen-Group A Beta Hemolytic Strep
hazard. - Culture on Todd Hewitt Broth for Fluorescence microscopy of beta
- Accidents or injuries must be reported immediately. Strep
- Nasopharyngeal swab is the specimen of choice to detect carrier
GENERAL GUIDELINES state of N. meningitidis and to detect presence of B. pertusis and H.
1. The specimen should be from the infection site and not influenza
contaminated by the surrounding area (e.g., culture within a
wound and not the surface or the surrounding skin). SPUTUM
2. Whenever possible, the specimen should be collected before - May often be contaminated with normal flora so it is important to
antimicrobials are administered. evaluate the quality of specimen
3. Appropriate collection devices and containers should be used and - Note the number of Squamous epithelial cells/LPF and PMN’s to
must be sterile. Aseptic technique is required. evaluate acceptability of specimen
4. The specimen container should be labeled with the patient’s - Bartlett’s classification
identification, the date and time of collection, and the source of - A gram stain is performed on all sputum sample; collected ideally
specimen. in AM when its most concentrated
SPECIMEN PROCESSING URINE
SPECIMEN TRANSPORT - Clean catch mid-stream- specimen of choice for bacterial culture
- Ideally specimens should be transported to the lab within 2hrs of - Catheterized- for those unable to urinate
collection - Suprapubic urine- anaerobic culture
- Must be preserved or refrigerated if not processed
SPECIMEN PRESERVATION - Major Cause of UTI
Boric acid – added to urine to maintain accurate colony count - Causes UTI among young sexually active females
0.025% SPS – anticoagulant for blood culture - MAC and BAP are suitable combination for culture
Heparin – for viral culture and can inhibit gram (+) organisms - Colony count should be performed on all urine samples
Note: EDTA and Citrate – not to be used in Microbiology
SPECIMEN STORAGE
CSF – should be kept at 37 degC/room temp.
Urine, Stool, Viral specimens
Sputum, Swab- Keep in refrigerator for 4 degC
Serum for serology – freeze -20 degC fo 7 days
Tissue specimens – for long term storage kept frozen for -70 degC
WAYS TO FACILITATE ANAEROBIC CULTIVATION
- Boiling culture medium to remove oxygen POUR PLATE METHOD
- Use of Gas-Pak Jar STOOL
- Anaerobic culture- are incubated at 35-37 degC - Detection of enteric pathogens
- Stool specimen not processed within 2hrs of collection should be
CLINICAL SPECIMENS placed on transport media Cary Blair
BLOOD - This specimen is routinely screened for Salmonella, Shigella and
- Avoid normal skin flora while collecting for blood culture Campylobacter
- Most common anticoagulant is sps - Maybe diretly plated on differential plate EMB or Mac and onto a
- Add 1% gelatin inhibits Neisseria, G. vaginalis, P. anaerobius so to selective media HEA or SSA and enrichment
counteract, Add 1% gelatin Dilution factor for blood to medium is
1:10 GENITAL TRACT SPECIMENS
- Amount to be collected Adult: 10ml Aerobic: 5ml Anaerobic: 5ml - To detect presence of N. gonorrhea, G. vaginalis and C.
- Children:1-5 ml trachomatis which is common cause of cervitis
- Routine blood culture 7 days
- Brucellosis: 3-4weeks • Leptospirosis: 8 weeks
CSF (CEREBROSPINAL FLUID)
- Usually collected using 3-4 tubes