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Respiratory Specimen Collection

Collection Procedure:

LTD-Respiratory: Specimen Collection  Version 8

Respiratory Specimen Collection

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)

1. Two flocked swabs should be collected if a rapid Ag test is ordered

and 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


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 08/2016

BioFire Package Insert. Respiratory FilmArray Verification Study CHMCA 2016

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