Www.testmenu.com



ASCENSION CRITTENTON HOSPITAL

PATHOLOGY DEPARTMENT POLICIES AND PROCEDURES

|Manual |MICROBIOLOGY |

|Procedure |Microbiology Specimen Collection |

PRINCIPLE

Specimen collection and transportation are critical considerations because the quality of a laboratory’s work may be limited by the nature of a specimen and its condition on arrival in the lab. The material to be cultured should be collected where the suspected organism is most likely to be found and with as little external contamination as possible. Whenever possible, specimens should be obtained before antimicrobial agents have been administered. Further, specimens should be of sufficient quantity to permit complete examination and transported to the lab as quickly as possible if the results of analyses are to be valid.

Specimen Transport

Ideally, all routine specimens are to be delivered to the laboratory within one hour of collection. All STAT specimens are to be delivered to the laboratory immediately after collection.

Specimen Storage

Transport or process all specimens immediately upon collection where possible. Individual specimen collection and storage criteria is available in the body of this document. If a specimen is collected and the source is not in this document, contact the lab. A tech will consult the various references located at the end of this section.

Specimen submitted for culture must be sent in a sterile, tightly sealed container. There must be no external spillage. Specimen containers are transported to the lab in disposable biohazard bags.

Specimen Labelling

Specimens must be labelled with the following:

1. Name of patient

2. Hospital number

3. Source of specimen

4. Time of specimen collection

5. Date of specimen collection

Requisition Information

1. Requisition forms must contain the following information:

a. Name of patient

b. Hospital number

c. Source of specimen

d. Time of specimen collection

e. Date of specimen collection

f. Name of physician

g. Test requested

2. All requisition slips will be timed in upon arrival at the lab and handed to a technologist.

Requisitions Required

Test requests on specimens are either ordered in the hospital’s computer system or another test request form. If a specific test is not listed in the computer test menu or any form, then use Miscellaneous 7230 form.

Computer ordered tests are ordered after the specimen has been collected. The source for each test requested is also entered in the computer. After the test(s) are ordered, computer labels will be generated that have all the required information as outlined above Requisition Information.

Unacceptable Specimens

1. Specimens will not be processed under the following conditions:

a. Visible evidence of hair or fecal contamination

b. Visible external container contaminated with blood, urine, or feces

c. Leaking container

d. Specimen on dry swab

e. 24 hour sputum for AFB

f. 24 hour urine for AFB

g. Patient ID on the requisition and that on the specimen differ

h. Incomplete or incorrect labelling

i. Incomplete or incorrect requisition slips

j. Specimens received in non-sterile containers (except stools)

k. Materials received in fixative

l. Foley catheters

m. Specimens received after prolonged delay

n. Insufficient amount of specimen

o. More than one specimen received from same site in one day for the same test without prior consultation with the physician (occult bloods are exempt)

p. Four or more blood cultures ordered in a 24 hour period without prior consultation with the physician

q. Fluids arriving in large, glass vacutainer bottles

2. If the laboratory receives an unacceptable specimen, Microbiology personnel will notify the floor and request a new specimen. The unacceptable specimen will be kept refrigerated until a new specimen is obtained. If a new specimen cannot be obtained, microbiology personnel will consult with the ordering physician before processing the specimen.

Specimen Collection

Specimens are collected in a variety of sterile containers as outlined under the various specimen headings. Occasionally, the use of a double swab culturette is indicated. The following are the instructions for the Copan culturette and Port-A-Cul tubes:

Copan Culturette Directions

a. Peel open sterile pack.

b. Remove swab and take specimen.

c. Remove and discard plug from the tube.

d. Insert swab into tube and push in plug.

e. Attach patient ID label and transport to lab in a transport bag.

Port-A-Cul Tubes

f. Using aseptic techniques, obtain a specimen with a swab.

g. Carefully loosen the screw cap and quickly insert the swab into the tube to within approximately 5 mm from the bottom of the medium.

h. Break the shaft of the swab evenly with the lip of the tube to allow easy access to the shaft.

i. Quickly replace and tighten the cap. Minimize exposure of the medium to the air.

j. Attach patient ID label and transport to the lab in a transport bag.

Abscess

Tissue or fluid is always superior to swab specimen. Sampling of surface area can introduce colonizing bacteria is not involved in infectious process.

1. Abscess, open

a. Remove surface exudate by wiping with sterile saline or 70% Ethanol.

b. Aspirate with a needle and syringe if possible, or pass culturette swabs deep into the lesion.

c. Firmly sample the lesion’s advancing edge.

d. If aspirated, remove needle and replace the original seal on the syringe. Alternately, inject the material into a sterile container.

e. If using swabs, place the swabs immediately in the culturette that has transport medium.

f. Transport to the lab within two hours.

2. Abscess, closed

a. Remove surface exudate by wiping with sterile saline or 70% Ethanol.

b. Aspirate abscess wall material with needle and syringe.

c. Remove the needle and replace the original seal on the syringe. Alternately, inject the material into a sterile container.

d. Transport to the lab within two hours.

Acid Fast Culture

1. Specimens are collected as outlined under specimen type.

2. The amounts for various specimen types are outlined below. Note that greater volumes will increase the chance of organism recovery.

a. CSF: 2 mL sterile screw cap tube

b. Respiratory: 1–10 mL

c. Sterile body fluids: >10 mL

d. Urine: >20 mL

e. Abscess: double swab culturette

3. Deliver the specimens to the lab within the time limits for specimen type.

Amniotic

1. Swabbing or aspiration of vaginal membrane is not acceptable because of vaginal contamination.

2. Aspirate via amniocentesis, cesarean section, or intrauterine catheter.

3. Transfer the fluid into a sterile screw-cap container.

4. Transport to the lab within 15 minutes at room temperature.

Anaerobic Culture

1. Specimens are collected as outlined under specimen type.

2. When using swabs, use the Port-A-Cul System in place of or in addition to the culturette.

3. For liquid or solid specimens, use a sterile screwcap tube.

4. Deliver the specimens to the lab within the time limits for specimen type.

Blood Cultures

1. MEDIA:

The following media are available for use with the BD BACTEC™ FX system:

BACTEC™ Standard/10 Aerobic /F

Recommended for 3.0–10.0 mL (8.0–10.0 mL optimal) blood volume.

BACTEC™ Plus Aerobic /F

Contains resins for antibiotic neutralization. Recommended for use in adult populations due to higher blood volume capacity and resins. Recommended for 3.0–10.0 mL (8.0–10.0 mL optimal) blood volume.

BACTEC™ Standard Anaerobic /F

Recommended for 3.0–7.0 mL (5.0–7.0 mL optimal) blood volume.

BACTEC™ Peds Plus

Contains resins for antibiotic neutralization.Optimized to detect organisms associated with pediatric septicemia and for low blood volumes (1.0–3.0 mL optimal; 0.5–5.0 mL recommended).

3. VOLUME:

The volume of blood cultured is critical because the number of organisms per millilimter of blood in most cases of bacteremia is low, especially if the patient is on anti-microbial therapy. In infants and children, the number of organisms per milliliter of blood during bacteremia is higher than adults, so less blood is required for culture.

Children: 1–5 mL of blood per venipuncture. Transfer the entire amount to a BACTEC™ Peds Plus/F vial.

Adult: 16–20 mL of blood per venipuncture. If it is impossible to draw the required amount, aliquot as follows:

Amount per Venipuncture Amount in BACTEC Plus Aerobic Vial Amount in BACTEC Plus Anaerobic Vial

16–20 mL Split equally between aerobic and anaerobic vials

13–16 mL 8 mL 5–8 mL

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 8–10 mL of blood (BACTEC Peds Plus/F: 1–3 mL). The use of lower or higher volumes may adversely affect recovery and/or detection times.

Technologist processing blood cultures must perform visual examination to check for adequate volume of blood specimen before loading on analyzer. If blood culture vials are not properly filled, communicate the issue to the draw person and document feedback on Microbiology/Serology Department Specimen Rejection Log indicating if specimen was accepted (e.g., volume low but patient was a difficult draw) or new specimen was requested.

3. SPECIMEN LABELING

Each vial should be labeled with the appropriate patient information:

Patient’s name.

Hospital number (Patient ID).

Patient’s location (room and bed #).

Date and time of collection.

Collector’s initials.

Description of draw site if obtained through a catheter or line.

Each draw set must be accompanied by a test order label bearing the patient’s unique specimen barcode and displaying all the information listed above.

4. NUMBER AND TIMING

Most cases of bacteremia are detected using two to three sets of separately col¬lected 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. Orders for more that three blood cultures within a 24 hour time span must be confirmed by contacting the ordering physician.

Bordetella pertussis Culture

1. Specimen requirements: Nasopharyngeal swab

2. Container: N/P culturette

3. Collection:

a. Shape the flexible swab into the contour of the nares and into the nasopharynx.

b. Pass the swab gently through the nose.

c. Leave swab in place near septum and floor of nose for 15–30 seconds.

d. Rotate and remove. Place swab in holder containing transport media.

4. Storage: Transport specimen to the laboratory immediately or inoculate medium immediately. Organism is extremely sensitive to drying.

Chlamydial Culture

1. Applies to: Cervical Chlamydia Culture, Eye Swab Chlamydia Culture, Respiratory Chlamydia Culture, and Urethral Chlamydia Culture.

2. Specimen Requirements: Chlamydia is intracellular. Obtain swab specimens containing columnar epithelial cells of conjunctiva, cervix, posterior nasopharynx, throat, rectum, or urethra.

3. Container: Specific Chlamydia holding transport medium and Dacron swabs must be used.

4. Collection

a. Urethra

Remove mucus/pus. The swab should be inserted 2–4 cm into the urethra. Use firm pressure to scrape cells from the mucosal surface. If possible repeat with second swab. Patient should not urinate within one hour prior to specimen collection.

b. Cervix:

Remove mucus/pus with a Dacron swab and use firm and rotating pressure to obtain specimen with another Dacron swab. May be combined with a urethral swab into same transport medium. This two-swab method is highly recommended.

c. Rectum:

Sample anal crypts with Dacron swab.

d. Conjunctiva:

Remove mucous and exudate. Use a Dacron swab and firm pressure to scrape away epithelial cells from upper and lower lids.

e. Posterior nasopharynx or throat:

Collect epithelial cells by using a Dacron swab.

5. Storage Instructions: Transport inoculated transport medium to the laboratory immediately. Specimens must be refrigerated if stored or transported within two days or frozen at −70°C if stored more than two days.

Corynebacterium diphtheria Culture

1. Applies to: Throat culture, nasopharyngeal culture, membrane culture.

2. Specimen Requirements: Throat swab, nasopharyngeal swab, membrane swab. Both a throat and N/P culture should be taken.

3. Container: Double swab culturette, N/P culturette.

4. Collection: Transport specimen to the lab immediately.

CSF

1. Collection is performed by physician.

2. Disinfect site with 2% iodine.

3. Insert needle with stylet at L3-L4, L4-L5, or L5-51 interspace.

4. Upon reaching subarachnoid space, remove stylet, and collect 1–2 mL of fluid into each of 3 or 4 sterile, leak-proof tubes.

5. Deliver to the lab within 15 minutes at room temperature.

Dental Culture

Includes: gingival, periodontal, periapical, Vincent’s Stomatitis.

1. Carefully cleanse gingival margin and supragingival tooth surface to remove saliva, debris, and plaque.

2. Using periodontal scaler, carefully remove subgingival lesion material.

3. Transfer the material to the swabs of culturette and/or Port-A-Cul tubes. Alternatively, pass the swabs over and into the lesions.

4. Place the swabs immediately in the cultures or Port-A-Cul tubes that have the transport medium.

5. Deliver to the lab.

Ear

1. Inner: Tympanocentesis is reserved for complicated, recurrent, or chronic persistent otitis media.

a. Collection is performed by the physician.

b. For intact eardrum, clean ear canal with soap solution, and collect fluid via syringe aspiration technique.

c. For ruptured ear drum, collect fluid on flexible-shaft swab via auditory speculum.

d. If aspirate or biopsy, use Port-A-Cul tubes and transport for ≤2 hours at room temperature for both aerobic and anaerobic cultures.

2. Outer

a. Use moistened swab to remove any debris or crust from ear canal.

b. Obtain sample by firmly rotating swab in outer canal.

c. For otitis externa, vigorous swabbing is required because surface swabbing may miss streptococcal cellulitis.

Eye

1. Conjunctiva

a. Sample eye by rolling N/P swab over conjunctiva.

b. Place swab in culturette with transport media and deliver to the lab.

2. Corneal scrapings

Take swabs for culture prior to anesthetic application; corneal scraping scan be obtained after.

a. Obtain conjunctival swab specimen as described above.

b. Instill two drops of local anesthetic.

c. Using sterile spatula, scrape ulcers or lesions, and inoculate scrapping directly into medium.

d. Apply remaining material to a clean glass slide for staining.

Feces

1. Routine

a. Do not routinely perform stool cultures for patients whose length of stay was more than three days and whose admitting diagnosis was not gastroenteritis.

b. Pass stool directly into clean, leak-proof wide-mouth container.

c. If available, transfer the stool to an enteric transport system.

2. Clostridium difficile toxin

a. Patient should be passing more than five stools with liquid or soft consistency per 24 hours. Soft stool is defined as stool assuming the shape of its container.

b. Pass stool directly into clean, leak-proof wide-mouth container.

3. Clostridium difficile culture

a. Pass stool directly into clean, leak-proof wide-mouth container.

b. Deliver to the lab immediately so that the stool can be transferred to a Port-A-Cul tube.

4. Leukocytes

a. Pass stool directly into clean, leak-proof wide-mouth container.

b. Specimen can be preserved indefinitely if transferred to 7.5% formalin and/or PVA.

5. Rectal Swab

a. Insert swab one inch beyond anal sphincter.

b. Rotate swab to sample anal crypts.

c. Feces should be evident on the swab.

6. Ova & Parasites

a. Pass stool directly into clean, leak-proof wide-mouth container.

b. If available, transfer stool to O & P transport system.

7. Occult Blood

a. Patient should not receive vitamin C for three days prior to testing. Antacids may cause false negative results.

b. Recommended for 72 hours prior to testing: High bulk, red meat free diet with restriction of peroxidase-rich vegetables (turnips, horseradish, artichokes, mushrooms, radishes, broccoli, bean sprouts, cauliflower, apples, oranges, bananas, cantaloupes, grapes). This will decrease incidence of false positives.

c. Also to be avoided are alcohol and aspirin (especially together), therapeutic iron, halogens and ametidine, and gastric irritants (steroids, all nonsteroidal anti-inflammatory drugs, colchicine).

d. Pass stool directly into clean, leak-proof wide-mouth container.

e. Submit stool or inoculate an occult-blood card. Follow the instructions accompanying the occult-blood card.

8. Rotavirus

a. Pass stool directly into clean, leak-proof wide-mouth container. Alternatively, collect a rectal swab.

b. A minimum of 10 g of stool or two rectal swabs is required.

Fluids

Includes abdominal, ascites, bile, joint pericardial peritoneal, pleural, synovial, dialysate, gallbladder, etc.

1. Disinfect overlying skin with 2% iodine tincture.

2. Obtain specimen via percutaneous needle aspiration or surgery.

3. Transfer fluid into a sterile, screwcap container, SPS or heparin tube or collect specimen with a culturette. Do not submit swab dipped in fluid.

4. If using a syringe and needle, remove the needle and replace the original seal on the syringe.

5. Transport immediately to the lab.

Fungus Culture

1. Specimens are collected as outlined under specimen type.

2. The amounts for the various specimen types are outlined below. Note that greater volumes will increase the chance of organism recovery.

a. CSF: 2 mL

b. Respiratory: 3–5 mL

c. Sterile body fluids: >10 mL

d. Urine: >20 mL

e. Abscess: Double swab culturette

3. Deliver the specimens to the lab within the time limits for specimen type.

Gastric Biopsy for H. pylori

1. Patient Preparation

Antibiotics and bismuth, when used suboptimal therapy, can suppress but not eliminate the organism. Therefore, the patient should not have used these agents for several weeks prior to the test. Following inadequate therapy, the organism may regrow in a patchy manner and may not be detected by random biopsies. The patient should not have received the proton pump inhibitor, omeprazole, prior to the test as this drug has been shown to have a bactericidal effect on the organism.

2. Biopsy Location

Because of the patchy nature of H. pylori infection, it is generally recommended that two specimens be obtained from each patient: one from the immediate prepyloric area and the second from the proximal antrum on the lesser curvature. Biopsies should be obtained from normal appearing mucosa as eroded or ulcerated mucosa may have fewer organisms.

3. Procedure

a. The tray with the hpfast agar should be evaluated before the test. The agar should have a light peach color. A very light green color may indicate contamination and the subsequent result should be interpreted cautiously. A tray with a dark green or blue agar color should be discarded. Allow the tray to reach room temperature before use.

b. Free the specially designed biopsy pick from the tray by squeezing the tray from both sides. The biopsy pick is designed at one end with a needle tip and spatula to remove the specimen from the forceps.

c. Peel back the label and place the specimen on the top of the agar. Use the reverse end of the biopsy pick to insert the specimen deep into the agar gel. The specimen should be exposed to as much agar as possible to enhance the likelihood of a positive reaction.

d. Transport the specimen to the laboratory within 15 minutes at room temperature.

Gastric wash or lavage fluid

Collect in early morning before patients eat and while they are still in bed.

1. Introduce nasogastric tube orally or nasally to stomach.

2. Recover sample and place in sterile screwcap container.

3. Before removing tube, release suction and clamp it.

4. Deliver to the lab within 15 minutes at room temperature or else neutralize within one hour of collection.

5. Specimens must be processed promptly because mycobacteria die rapidly in gastric washing. Neutralize each 35–50 mL of gastric washings with 1.5 mL of 40% anhydrous Na2HPO4.

Genital

1. Bartholin

a. Disinfect skin with 2% iodine tincture.

b. Aspirate fluid from ducts.

c. Transfer the fluid into a sterile screwcap container.

d. Transport to the lab within two hours at room temperature.

2. Cervix

Viral and chlamydial test require separate collection and transport kits. See viral culture or chlamydial culture.

a. Visualize cervix with speculum without lubricant.

b. Remove mucus and/or secretions from cervix with swab and discard swab.

c. Firmly yet gently, sample endocervical canal with sterile swab.

d. Place swab in culturette with transport media and transport to the lab within two hours at room temperature.

3. Cul-de-sac

a. Submit aspirate or fluid in a sterile screwcap tube.

b. Transport to the lab within two hours at room temperature.

4. Endometrium

a. Collect transcervical aspirate via telescoping catheter.

b. Transfer entire amount into a sterile screwcap container.

c. Transport to the lab within two hours at room temperature.

5. Urethra

a. Remove exudate from urethral orifice.

b. Collect discharge material on swab by massaging urethra against pubic symphysis through vagina.

c. If no discharge can be obtained, wash external urethra with betadine soap, and rinse with water. Then insert urethrogenital swab 2–4 cm into urethra, and rotate swab for two seconds.

d. Place swabs immediately in a culturette that has transport media and transport to the lab within two hours at room temperature.

6. Vagina

a. Wipe away excessive amount of secretion or discharge.

b. Obtain secretions from mucosal membrane of vaginal vault with swab.

c. Place swab immediately in a culturette that has transport media and transport to the lab within two hours at room temperature.

d. For intrauterine devices, place entire device into sterile container and submit at room temperature.

Hair

1. No cleaning of scalp is needed.

2. Select infected areas, and with forceps, epilate at least 10 hairs.

3. For hairs broken off at the scalp level, use a scalpel or a blade knife.

4. Place fairs in a clean or sterile container.

Influenza A & B Antigen

1. Specimen requirements: The following specimens are appropriate: Nasal specimens collected on the appropriate dry CultureSwab.

2. Collection: Collect appropriate dry swab as usual for Nasal specimen. Do not place swabs or specimen in viral transport medium. Transport to lab within two hours at room temperature.

Legionella Culture

1. Applies to: Lung tissue, legionella culture; tissue, legionella culture; pleural fluid, legionella culture; transtracheal aspiration, legionella culture.

2. Specimen Requirements: Lung tissue, other body tissue, pleural fluid, other body fluid, transtracheal aspiration, bronchoalveolar lavage, and bronchial brushing.

3. Container: Sterile screwcap container.

4. Collection: Transport specimen to the laboratory within two hours at room temperature.

Mycoplasma Culture

1. Specimen Requirements: The following specimens are appropriate: Throat swabs (send two swabs), sputum, bronchial washings, tracheal aspiration, cerebrospinal fluid, heparinized blood.

2. Collection: Transport specimen to the la within two hours at room temperature. Do not place specimens in preservative. Do not use swabs with wooden sticks.

Nail

1. Clean nail with 70% alcohol.

2. For a specimen of the dorsal plate, scrape the outer surface and discard the scrapings. Then scrape the deeper portion for a specimen.

3. Remove a portion of debris from under the nail with a scalpel.

4. Collect the whole nail of nail clippings.

5. Place all material in a clean or sterile container.

Respiratory Tract, upper

1. Oral

a. Remove oral secretions or debris from surface of lesion with swab and discard swab.

b. Using second swab, vigorously sample lesion, avoiding any areas of normal tissue.

c. Place swab in culturette and transport to the lab within two hours at room temperature.

2. Nasal

a. Use swab premoistened with sterile saline.

b. Rotate swab against nasal mucosa.

c. Place swab in culturette and transport to the lab within two hours at room temperature.

3. Nasopharynx

a. Gently insert Dacron N/P swab into posterior nasopharynx via nose.

b. Rotate swab slowly for five seconds to absorb secretions.

c. Remove swab, place swab in culturette and transport to the lab within two hours at room temperature.

4. Throat

a. Depress tongue with tongue depressor.

b. Sample posterior pharynx, tonsils, and inflamed areas with sterile swab.

c. Place swab in culturette and transport to the lab within two hours at room temperature.

d. Throat cultures are contraindicated for patient with inflamed epiglottis.

Respiratory Tract, lower

1. BAL, BBW, tracheal aspirate

a. Place aspirate or washing into sputum trap.

b. Place brush into sterile container. Do not add saline.

c. Submit specimen to the lab within two hours at room temperature.

2. Sputum, expectorate

a. Collect specimen under direct supervision of nurse or physician.

b. Have patient rinse or gargle with water.

c. Instruct patient to cough deeply to produce lower respiratory specimen (not postnasal fluid).

d. Collect specimen into sterile container.

e. For patients unable to produce specimen, respiratory therapist should collect via suction.

f. Submit specimen to the lab within two hours at room temperature.

3. Sputum, induced

a. Have patient rinse mouth with water after brushing gums and tongue.

b. With aid of nebulizer, have patient inhale approximately 25 mL of 3–10% sterile saline.

c. Collect induced sputum into sterile container.

d. Submit specimen to the lab within two hours at room temperature.

RSV – Respiratory Syncytial Virus

1. Collect both an N/P and a throat specimen as outlined above.

2. Nasal washings is the specimen of choice and is collected as follows:

a. Wipe excess mucus from nose with facial tissue.

b. Empty 5 cc sterile saline into clean cup.

c. Suction fluid into bulb syringe.

d. Have patient lying down with head tipped back.

e. Insert bulb into nostril until opening is occluded.

f. In one motion, gently squeeze fluid into nostril and then release to collect fluid again into bulb.

g. Empty material into a sterile container and transport to the laboratory immediately.

Skin dermatophytosis

1. Cleanse affected area with 70% alcohol.

2. Gently scrape surface of the skin at active margin of lesion. Do not draw blood.

3. Collect enough scrapings to cover the head of a thumb nail.

4. Place sample in clean container.

5. If specimen is submitted between glass slides, tape slide together and submit in envelope.

6. Submit the specimen to the lab within 24 hours at room temperature.

Tissue

1. Submit in sterile screwcap container.

2. For small samples, add several drops of sterile saline to keep moist. Do not allow tissue to dry out.

3. Always submit as much tissue as possible.

4. Never submit swab that has simply been rubbed over surface.

5. Transport to lab within 15 minutes at room temperature.

Urine

1. Female, midstream

a. Open packet of benzalkonium chloride antiseptic.

b. While seated on the toilet, spread labia majora (outer folds).

c. With the first towelette, wipe one side of the labia minor (inner fold) using a single downward stroke. Discard towelette.

d. With the second towelette, repeat procedure on opposite side using a single downward stroke. Discard towelette.

e. With third towelette, cleanse meatus (center area) with a single downward stroke. Discard towelette.

f. First void in toilet. After several milliliters have passed, bring collection cup into “midstream” to collect urine specimen without stopping flow of urine.

g. Screw cap onto specimen container.

h. Transport specimen to the lab within two hours at room temperature.

2. Male

a. Open packet of three towelettes.

b. Retract foreskin, if present.

c. With the first towelette, cleanse the meatal orifice with a single downward stroke. Discard towelette.

d. Repeat Step 3 with the two remaining towelettes.

e. First void in toilet. After several mL have passed, bring collection cup into “midstream” to collect urine specimen without stopping flow of urine.

f. Screw cap onto specimen container.

g. Transport specimen to the lab within two hours at room temperature.

3. Straight Catheter

a. Thoroughly clean urethral area with soap and water.

b. Rinse area with wet gauze pads.

c. Aseptically insert catheter into bladder.

d. Allow approximately 15 mL to pass, and then collect urine to be submitted in sterile screw cap container.

e. Transport the specimen to the laboratory within two hours at room temperature.

f. Not recommended for routine urine culture because of potential contamination problems. Procedure may introduce urethral flora into bladder.

4. Indwelling Catheter

a. Disinfect catheter collection port with 70% alcohol.

b. Use needle and syringe to aseptically collect 5–10 mL of urine.

c. Transfer sample to sterile screwcap container.

d. Transport the specimen to the laboratory within two hours at room temperature.

Viral Culture

1. Applies to: Adenovirus Culture; cytomegalovirus culture; enterovirus culture; Epstein-Barr virus culture; Herpes Simplex virus culture; influenza virus culture; parainfluenza virus culture; respiratory syncytial virus culture; varicella-Zoster virus culture. Indicate on the request the type of infection and/or virus expected.

2. Specimen Requirements: Blood, CSF, dermal, ocular, genital, mucosal, oral, rectal, respiratory, stool, tissue, urine, biopsy.

3. Volume: 1 mL fluid or one swab. Minimum is 0.5 mL fluid or 0.5 g tissue and stool.

4. Container: Viral transport medium for swabs; sterile screwcap tube or container for fluids, feces, nasal washings, urine or biopsy (no preservative); lavender top tube for blood, bone marrow, and buddy coat.

5. Collection: Always consult the laboratory for specific details prior to collecting specimen. Specimen should be collected during the acute phase of the disease as follows:

a. Throat swab: Carefully rub the posterior nasopharyngeal wall with a dry, sterile swab. Avoid touching tongue or buccal mucosa. Use viral transport.

b. Blood: Collect 5 mL whole blood into lavender top (EDTA) tube. Consult laboratory for specific recommendations.

c. Feces: Collect 4–8 g feces (about the size of a thumbnail), and place in a clean, leak-proof container. Do not dilute the specimen (into virus transport medium) or use preservatives.

d. Rectal Swab: Insert a sterile swab 2–4 inches into the rectum and rub the mucosa. Use viral transport.

e. Cerebrospinal fluid: Collect 1 mL CSF aseptically in a sterile dry screwcap vial. Do not use viral transport medium.

f. Skin lesions: Open vesicle and absorb exudate into a dry swab, and/or vigorously scrape base of freshly exposed lesion with a swab to obtain cells that contain viruses. If enough vesicle fluid is available, aspirate the fluid with a fine gauge needle and tuberculin syringe, and place the fluid into cold viral transport medium.

g. Urine: Collect clean-catch, midstream urine in a leak-proof, sterile, plastic container. Do not use viral transport medium.

h. Tissue: Use a fresh set of sterile instruments to collect each tissue. Place each specimen in a separate dry, sterile nontoxic leak-proof container. Identify each tissue with patient’s name, type of tissue, and date collected. The tissue should eventually be placed in viral transport media.

i. Eye swab or scraping: Use a swab to collect conjunctival material or take conjunctival scrapings with a fine sterile spatula and transfer the scraping to a viral transport medium.

j. Genital swab: See skin

Catheterized Specimen

1. Free flow from the catheter mouth is the ideal specimen. (Urine from collection bag is generally unsuitable.) Approximately 5–10 mL are sufficient.

Urine collected for indwelling Foley catheter tips are unsuitable for culture (Catheter tips are invariably contaminated with urethral organisms).

2. If urine specimens for culture cannot be brought to the Laboratory within 30 minutes of collection, the specimen must be refrigerated.

For collection of specimens not listed above or for unusual test requests, please contact the Microbiology Lab.

REFERENCES

1. Barron, E.J. and Finegold, S.M., Baily and Scott’s Diagnostic Microbiology, 9th ed., St. Louis: C.V. Mosby Co.; pages 4, 54, 178, 1994.

2. Balows, A., Hausler, W.J., Herrmann, K.L., Isenberg, H.D., and Shadomy, H.J., Manual of Clinical Microbiology, 5th ed., Washington, D.C.: American Society for Microbiology; pages 14–30, 306, 513, 588–590, 602–603, 633–634, 644, 660, 678, and 694.

3. Miller, J.M., and Wentworth, B.B., Methods for Quality Control in Diagnostic Microbiology, Washington D.C.: American Public Health Association; pages 18–20, 23–26, 30, 33, 35–41, 50–52, 1985.

4. Isenberg, H.D., Schoenknecht, F.D., and von Graevenitz, K., Cumitech 9: Collection and Processing of Bacteriological Specimens, Washington, D.C.: American Society for Microbiology; pages 3–4, 1979.

5. BBL Port-A-Cul Specimen Collection and Transport Products Package Insert. Cockeysville, MD; BBL Microbiology Systems; Revised Feb 1993.

6. Bartlett, R.C., Ellner, P.D., and Washington, J.A., Cumitech 1: Blood Cultures: American Society for Microbiology; page 2, 1974.

7. Hpfast Package Insert, GI Supply, A Division of Chek/Med Systems, Inc., Camp Hill, PA, 1995.

8. Buck, G.E. Campylobacter pylori: A New Organism Implicated as a Cause of Gastritis and Peptic Ulcers. Clinical Microbiology newsletter. Vol. 19, No. 18, pages 141–144, 1987.

9. Bactec Package Inserts PP-105 (Standard/10 Aerobic/F Culture Vials) 6/94, and MA-0034 (PEDS Plus/F Culture Vials) 2/94, Becton Dickinson Diagnostic Instrument Systems, Sparks, MD. MA-00344 (Standard Anaerobic/F Culture Vials). 5/93.

10. Howard, B.J., Klaas, J., Ruben, S.J., Weisseld, A.S., Tilton, R.C., Clinical and Pathogenic Microbiology, St. Louis: C.V. Mosby Co.; pages 208, 223–225, 1987.

11. Microbiology Inspection Checklist, College of American Pathologists, Northfield, IL, 1994 ed., question 0.4.1400 – 0.4.1425.

12. Becton Dickinson Directigen RSV Package insert, Cockeysville, MD. BD Microbiology Systems, Revised July 1991.

13. Copan Venturi Transystem Transport Swabs Package, Copan Diagnostic, Inc. New York, NY.

14. Isenberg, Henry D., Clinical Microbiology Procedures Handbook, American Society for Microbiology, Washington, D.C., pages 6.1.1–6.1.3, 6.6.1–6.1.5, 11.1.1–11.7.1, 1.1.13, 1.1.23–1.1.25, 1990.

15. Wampole Laboratories, Cranbury, NJ, Wampole Isolator 1.5 Microbial Tubes Package Insert, April 1993.

16. Laboratory Services Manual, DMC University Laboratories, pages 78, 84, 135, 145, 189, 1995.

................
................

In order to avoid copyright disputes, this page is only a partial summary.

Google Online Preview   Download