This standard operating procedure outlines the process for collecting and transporting sputum specimens from Sebbi Hospital to their laboratory. It details tasks and responsibilities, equipment needed, and collection procedures both when supervised by lab staff and when collected by the patient at home. The goal is to obtain a quality sputum sample that provides accurate test results by collecting thick, mucoid material from the lower respiratory tract, not saliva or other secretions, following biosafety precautions. Specimens must be at least 1mL in volume and transported refrigerated to the lab for analysis or referral within required timeframes.

1. Sputum Collection SOP

2. Objective and scope
This is a standard operating procedure (SOP) that outlines standardized methods for the collection and
transport of sputum specimens to and from the SH laboratory to ensure a quality specimen that provides
quality results. This SOP applies to staff of Sebbi Hospital and sputum samples processed with Sebbi Hospital
Lab
Abbreviations and definitions
Tasks and responsibilities
Tasks
Authorized Responsible
Review and update this procedure Lab Manager
Maintenance and control of the scientific content of this
SOP
Quality focal team Lab manager
Implementation of this SOP and ensuring that all
appropriate personnel are trained
Quality focal team Lab Manager
Reading and understanding this SOP prior to performing the
procedures described
All lab staff
Understanding and adhering to the patient preparation and
specimen collection components
All lab staff
Approve this SOP for use Medical Director
Equipment, Consumables, and PPE
Equipment,Consumables,andPPE
StrongCarbolFuchsin(Primarystain) Lenspaperandlenscleaningsolution
25%sulphuricacid(Decolorizingagent) Immersionoil
3%MethyleneBlue(CounterStain) Newslidesandnewslideholder
LightMicroscope Slidestoragebox
Spiritlamp Diamondpencil

3. Sputum Collection Procedures
The ideal sputum specimen is produced by repeated deep inhalation and exhalation of breath followed by a
cough as deep within the chest cavity as is possible for the patient. Sputum should consist of thick, mucoid,
white-yellow, sometimes blood-tinged, material from the lower airways and lungs (not saliva or oral/nasal
secretion). Collection of early morning specimens is preferred due to overnight accumulation of secretions.
However, specimens may be collected at any time from patients who have a deep cough that is readily
productive.
Supervised collection by Lab staff
1. When collected within the hospital, the phlebotomist should stand within viewing distance of the patient
during the procedure to provide assistance as needed; and to ensure that he/she is isolated from others
until sputum collection is complete.
2. Specimens should be collected in a well-ventilated area. The lab staff at phlebotomy assisting in collecting
the sputum, MUST observe the appropriate infection control precautions, i.e. wear a N95 mask and wear
gloves when in hand contact with blood or other potentially infectious materials anticipated
3. Collect sputum specimen in a sterile disposable, wide-mouth container
4. Prior to collection, label the specimen container with the patient lab number and the test required. Mark
the container ’1’ to differentiate it from the sputum collected at home - ( marked
’2’)
’2’)
5. Inform the patient that saliva and upper respiratory/nasal secretions are not sputum and are not
acceptable specimens
6. Instruct the patient to:
(a) Thoroughly clean his/her hands with soap and water. direct the patient to the hand washing station
with soap and a drier
(b) Breathe deeply a number of times and then cough from deep down within the lungs.
(c) Lean forward, breathe in and out slowly twice, hold breath for 2-3 seconds each time, and on third
time forcefully cough to bring up the sputum.
(d) Collect the sputum in the sterile container provided and avoid touching the inside or edge of the
specimen container or lid with their fingers
(e) Once collection has been completed, thoroughly clean his/her hands with soap and water.
7. Repeat the above sequence until an adequate amount of sputum is collected. This may take up to 1 hour.
If the patient is unable to produce enough sputum within 1 hour, decide if the patient is ?unable to
expectorate?, requires rescheduling for another attempt at collection, or needs to undergo sputum
induction.
8. Tighten the lid/cap on the container/tube and to avoid leakage
9. Estimate the volume of sputum collected by comparison with container/tube with markings. NOTE: As
a rule, a minimum volume of 1mL of sputum must be collected
10. After the specimen is collected, place specimen container in refrigerator or unless it is being analyzed
within 1 hour.
11. If the sputum is being referred out for further testing, it MUST be packaged in sealed zip lock bags with
sufficient absorbent material and marked with appropriate biohazard labeling. Complete all the relevant
fields on the Specimen Request Form accompanying the sample

4. Collection by patient at home
1. Provide the patient with a labeled specimen container with the appropriate identifying
information.Container marked ’2’ to indicate the specimen was collected at home
2. Provide the patient with a zip lock storage bag and absorbent material.
3. Inform the patient that saliva and upper respiratory/nasal secretions are not sputum and are not
acceptable specimens.
4. Instruct the patient to:
(a) Collect the sputum after getting out of bed, before the morning meal, and prior to taking
any medications.
(b) Collect the sputum in a well-ventilated area such as by an opened window or outside (c)
Thoroughly clean his/her hands with soap and water.
(d) Rinse his/her mouth with bottled or boiled water prior to collection of sputum.
(e) Breathe deeply a number of times and then cough from deep down within the lungs
(f) Lean forward, breathe in and out slowly twice, hold breath for 2-3 seconds each time, and on third
time forcefully cough to bring up the sputum.
(g) Collect the sputum in the sterile container provided and avoid touching the inside or edge of the
specimen container or lid with their fingers. item Replace the lid/cap after collection and close
tightly to avoid leakage.
(d) Once collection has been completed, thoroughly clean his/her hands with soap and water.
(e) Store the container in a zip lock bag with absorbent material. Bring the specimen container to the
hospital as soon as possible
1. Suggest placing thewater bottle/containerand specimen containerina placethat will remind the patient
to collect the specimen first thing in the morning upon rising.
2. Inform the patient that they must write down the time of collection on the specimen container or bring
this information when he/she brings back the specimen.
Receipt of Specimen Container
(a) Ask the patient at what time he/she collected the specimen and record their response on the container
(if not already present).
(b) Estimate the volume of sputum collected by comparison with container/tube with markings. If specimen
volume is inadequate (less than 1mL), try to collect another specimen from the patient while at the
hospital
(c) Refrigerate the specimen until transported to the laboratory. item Complete all the relevant fields on the
Specimen Request Form

5. Related documents and forms
(a) Specimen transportation SOP ( SH-LAB-P 06V01)
(b) Specimen request form ( SH-LAB-F 01V01)
Symptoms of infection with pathogens worked with in the laboratory ( SH-LAB-P 05V01