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Respiratory Specimen Collection

Collection Procedure:


Respiratory Specimen Collection

Policy Tech: Version 7

A. Nasal culture

Equipment: ESwab, mini-tip collection kit BD 220246

Insert the swab into a nare until resistance is met at the level of the
turbinate. Rotate the swab against nasal mucosa. Break swab off into the
transport tube containing 1.0mL of liquid media, send to laboratory.

B. Nasal Wash
(For Rapid RSV or Rapid Influenza virus, Adenovirus, or parainfluenza
virus antigen detection or viral culture).

Equipment: Sterile normal saline (NS) for irrigation, luki-trap, suction
catheter, gloves, M4 (VIRAL TRANSPORT MEDIA- VTM) medium.

1. Have the patient clear any nasal congestion before obtaining
    the specimen by blowing his nose or by nasal suctioning.

2. Connect the suction catheter to the luki-trap and then connect
    the luki-trap to wall suction.

3. "Wash" for nasal cells by irrigating both nares with 0.5 ml normal
     saline. Suction each nares to obtain cells not mucus.

4. Rinse the suction catheter with 1.0-1.5ml normal saline which
    helps move the specimen from the suction catheter into the lukitrap.

a. Place 0.5ml of saline wash into M4 (VIRAL
TRANSPORT MEDIA- VTM) medium and send both the nasal
wash and dilute M4 (VIRAL TRANSPORT MEDIA- VTM)
specimens to the lab.

b. If using a bulb syringe, empty contents of the bulb syringe
directly into sterile container. Place 0.5ml into M4 medium
for back-up culture and send both the nasal wash and the
M4 diluted specimen on ice to lab for testing.

NOTE: Do not add M4 viral medium to the specimen. The M4
(VIRAL TRANSPORT MEDIA- VTM) must be sent on ice. Send
immediately to Laboratory.

Special Instructions: The saline wash specimen is used for the direct
antigen assay. A portion of the specimen must be placed in M4 (VIRAL
TRANSPORT MEDIA- VTM) as it protects the viability of the virus for a
back up viral culture should the direct antigen assay be negative.


D. Nasopharyngeal (NPH)

Rapid RSV, Rapid Influenza, Adenovirus, Parainfluenza virus
1 or2 NPH flocked swabs BD 220251, M4 Remel 12520 (VIRAL

NPH culture (for N. meningitis or other bacteria)
ESwab minitip collection kit (green capped) BD 220246

NPH for Bordetella pertussis
1 NPH flocked swab BD 220251or NPH wire swab (green capped) BD
Send Flocked swab and place in M4 (VIRAL TRANSPORT MEDIA- VTM)

Specimen Collection (NPH culture and/or B. pertussis):
1. Hold the child securely. Remove a swab from the culturette. Slide the
    swab into the nare to the posterior nasopharynx.
2. Leave swab in place 20-30 seconds or as tolerated. Withdraw the
3. Repeat the process on the other nare with a second swab. Replace the
   swab into the culturette. For pertussis PCR, only one swab is used to 
   culture both nares.

Specimen Collection (Rapid RSV, Rapid Influenza, Adenovirus and
Parainfluenza virus, Respiratory FilmArray)
Two flocked swabs should be collected if a rapid Ag test is ordered
a Respiratory Virus isolation is ordered also. One swab is
    sufficient for a rapid test only
2. Slide the flocked swab into the nares to the posterior nasopharynx (at
    base of throat). Rotate the swab and allow 5-10 seconds for liquid to
3. The first swab can be placed back into the culturette and sent for rapid
4. Repeat collection with the second flocked swab. The second flocked
   swab is broken off into M4 (VIRAL TRANSPORT MEDIA- VTM). This
   specimen will be used for the back-up respiratory virus isolation culture
   if the rapid test is negative.

5. For FilmArray only one swab is necessary and it must be placed in M4
   transport medium.

E. Sputum

Equipment: Sterile cup (clear plastic, screw cap)

a. Collect specimen resulting from a deep cough into sterile cup.
Notify the physician if unable to obtain specimen.
b. A specimen may be obtained by sterile suction technique using
a Luki-trap by passing a sterile suction catheter along floor of
nose to nasopharynx. When the patient coughs, suction the
specimen into trap. Obtain specimen by suction only with
physician order.
c. Twenty-four hour sputum collections are not recommended for
culture. If possible, have the patient rinse mouth and gargle with
water prior to sputum collection. Instruct the patient not to spit
saliva or postnasal discharge into the container.
A gram stain smear result on sputum specimens will show
epithelial cells. A sputum specimen containing > 25 epithelial
cells per low power field has been contaminated with
oropharyngeal secretions during collection, indicating a poor
quality specimen for culture. Lab may indicate the need for
specimen recollection. Lab evaluates the specimen for the
predominant pathogenic morphotype if the physician requests a
culture on available specimen.
d. Sputum (acid fast bacilli or mycobacteria) First morning sputum
samples of 3.0-5.0 mL is optimal for possible recovery of
mycobacterial organisms. Acid fast stain performed on all
sputum specimens.

F. Tracheostomy Culture

Equipment: Luki trap, one pair sterile gloves, sterile suction
catheter, sterile normal saline for irrigation, Luki trap tubes,
disposable clean graduated cup, audit trail and label.

1. Connect suction catheter to rubber tubing side on
aspirating trap and connect the other side of
aspirating trap to the tubing from wall suction.

2. Draw up the appropriate amount of preservative free
sterile saline for irrigation into a syringe.

Under 1 year: 0.5ml;
1-3 years: 1ml;
3 years and older: 2ml.

Pour the remaining normal saline for irrigation into
graduated cup.

3. Suction the patient by putting on sterile gloves,
instilling saline from syringe (without needle),
suction patient while keeping the aspirating trap in a
vertical position.

4. If the specimen is in the catheter, suction a small
amount of normal saline for irrigation from
graduated cup to move secretions into aspirating
trap. Disconnect aspirating trap from suction tubing
and catheter. Connect tubing on aspirating trap to
other side of the trap.

5. Specimen should not be transported in the tube
system. Rupture of trap due to pressure changes
while in the tube system can occur.

G. Throat (Pharyngeal)


Tongue depressor

Group A antigen test (rapid) and culture: Rayon dual swab

Throat culture for GC only: Eswab collection kit

Routine culture (Haemophilus influenza, Strep pneumoniae):

ESwab collection kit BD 220245 or 220246 mini swab
NOTE: Do not obtain throat cultures if the epiglottis is
inflamed. Sampling may cause serious respiratory

1. Remove the swab from the collection kit package.
2. Depress tongue gently with tongue depressor.
3. Visualize throat area for obvious purulent areas. Swab between the 
   tonsillar pillars and behind the uvula. Avoid touching the cheeks,
   tongue, uvula or lips.
4. Swab back and forth across the posterior pharynx, tonsillar area
  and any inflamed or ulcerated areas to obtain sample.


For Group A Strep antigen test and Strep culture: Replace the two swab
specimens in the culturette.


By Laboratory Policy a Group A Strep Culture will be ordered and set up
following a negative Strep antigen test if a second throat swab is received by the

When group A Strep antigen (rapid) test and culture are ordered, the antigen
(rapid) test will be performed first. If the result is positive, the culture will not be
processed and the patient will not be charged for the culture. .

For throat GC culture and routine culture: the swab is broken off into the
transport tube containing 1.0mL liquid media,and sent to the lab.

Labeling: Label all specimen with at least 2 specimen identifiers, such as name,
medical record number, and date of birth



Bowden, V.R., Smith Greenberg, C.(2008). Pediatric Nursing Procedures, 2nd Ed.

Philadelphia: Wolters Kluwer/Lippincott Williams & Wilkins.

Isenberg, H.D. (2004). Clinical Microbiology Procedures Handbook, Volume 2.

Washington, D.C.: The ASM Press.

E swab Package insert. BD 220245 or BD 220246 mini Eswab. Becton

Dickenson 2010

BioFire Package Insert. Respiratory FilmArray Verification Study CHMCA 2016