COLLECTION AND TRANSPORT OF CLINICAL SPECIMENS

1.THROAT SWAB:
Indicated in : Sore throat
Technique of collection :
1.Patient’s head has to be tilted back and the throat well illuminated, depress the
tongue to see throat.
2. Rub the swab up and down on the throat and on any white patches in the tonsillar
area. Avoid the tongue and the cheeks.
3. Replace the swab in the transport tube. In case of Virus/Chlamydia/ Mycoplasma,
break off and leave the tip in tube.
4. Seal tube tightly and label with patient details.
5. Send the specimen to the laboratory with filled -in request form.
Commonly isolated pathogens:
• Streptococcus species
• Corynebacterium diphtheriae (calcium alginate swab)
• Hemophilus influenzae.

2. NASOPHARYNGEAL SWAB
1.Procured the swab and universal transport medium and check for expiry dates
2. Explained procedure to patient.
3.Hand hygiene to be done and put on the gloves and facemask
4.Keep the patient in a high-fowler’s position in bed with the back of the head
supported.
5.Place finger on tip of the nose and introduce a flexible swab for 5-6 cms with a slow,
steady motion along the floor of the nose. Once resistance is met rotate the swab
several times and withdraw the swab.
6. Break off top of swab, place in transport medium.
7.Remove personal protective equipment and label the container with necessary details.
8. Wash the hands
Commonly done for isolation and antigen detection in case of………
• Influenzae virus
• Respiratory syncytial virus
• Covid 19

2.SPUTUM
Indicated in : Lobar pneumonia , Broncho pneumonia, Atypical pneumonia,
pulmonary tuberculosis
Technique of Collection:
1.The best time to collect a specimen is in the morning, just after rising.
2.Sputum should be collected in a separate place or in area with negative pressure.
3.Rinse and spit with water to ensure there won’t be mouth bacteria in the sputum
collected.
4.Ask the patient to take a few deep breaths while pressing their hand lightly on their
stomach.
5.Ask to cough deeply, do not take shallow coughs from the throat or chest.
 6. Once the sputum (phlegm) is in the mouth to be released into a wide mouthed, screw
capped container.
7.Once the child starts to cough, sputum is obtained by suctioning through the
nasopharynx
8.Label the sample container, fill-up the requisition form properly and send it to
laboratory as early as possible
Processing :Gram staining, Z-N staining
Culture on Blood agar, chocolate agar, L-J medium(if suspecting TB)
Commonly isolated pathogens
▪ Streptococcus pneumoniae
▪ Hemophilus influenzae
▪ Staph aureus
▪ Pseudomonas aeruginosa
▪ Klebsiella pneumoniae

3.PUS:
Indicated in: Infected wound
Sample: Aspirated material for abscess, swab from surgical site infections.
Technique of collection:
Levine Method for swab culture
1. Cleanse wound with normal saline.
2. Remove/debride nonviable tissue.
3. Wait 2-5 minutes.
4. If ulcer is dry, moisten swab with sterile normal saline.
5. Culture the healthiest looking tissue in the wound bed.
6. Do not culture exudate, pus, eschar or heavy fibrous tissue.
7. Rotate the end of the sterile alginate-tipped applicator over a 1cm2 area for 5
seconds in zig-jag pattern across the wound.
8. Apply sufficient pressure to swab to cause tissue fluid to be expressed.
9. Use sterile technique to break tip of swab into collection device designed for
quantitative cultures.
10. Requisition form has to be filled in properly then send to the laboratory.
Commonly isolated pathogens
• Staph.aureus,
• Strep. pyogenes,
• Enterococci,
• Klebsiella spps
• Pseudomonas

4. VAGINAL SWAB :
Indicated in: Bacterial vaginosis, Trichomoniasis, Candidiasis
Technique of collection :
1.Hold swab in your right hand, placing thumb and forefinger in the middle of
the swab shaft.
2.Open the vagina with left index finger and thumb.
 3.Carefully insert swab into the inside opening of the vagina, about two inches, and
gently rotate swab for 10 to 30 seconds.
4.Make sure swab touches the walls of the vagina so that discharges is absorbed
by swab.
5.Unscrew the swab container with left index finger and thumb, then open it by using
right hand.
6.Keep the swab inside the container and close it tightly.
7.Ensure the proper labelling and transport it to the laboratory
Processing :
A wet mount is prepared using drop of saline (Parasitic trophozoites)
Gram stain is done
Culture : Blood agar, Mac Conkey’s agar, Chocolate agar.
Commonly identified /isolated pathogens
• Trichomonas vaginalis
• Gardnerella vaginalis
• Candida albicans

5. URINE:
Indicated in :
Bacterial UTI, Urethritis, Acute pyelitis, Prostatitis.
Technique of collection:
Types of specimens may be collected:
• First morning specimen
• 24 hours urine specimen
• Specimen from an indwelling catheter not from Uri bags
• Clean-catch midstream urine sample to be collected for culture
1. While collecting urine specimen, it is always important for the nurse to observe specific
protocols. Single random specimens may be taken at any time of the day or night. A 24-
hour urine specimen is an extremely important to know the changes that occur in the
kidneys.
2.The perineal area in women or the end of the penis in men should be cleaned with sterile
distilled water before the urine is collected.
3.Wiping the genitalia with a sterile wipe may stimulate the voiding reflex in infants. Urine
specimens need to be examined within 2 hours.
4.Don’t collect the urine sample from collection bags
5.Catheter puncture site must be sterilized before introducing the needle
6.At least 10 ml of urine midstream urine should be collected in a wide mouthed, screw
capped sterile container.
7.Container has to be properly labelled and to be sent to laboratory with filled in form.
8.A urine specimen should be refrigerated if it not sent to the laboratory within 2 hours.
Processing :
Microscopic examination – Saline wet mount, Gram stain
Chemical method: Triphenyl tetrazoline chloride test.
Enzymatic test; glucose oxidase.
Culture on: Nutrient agar, Blood agar, Mac Conkey’s agar.
 Commonly isolated pathogens:
• Escherichia coli,
• Klebsiella,
• Enterococci,
• Pseudomonas
• Citrobacter species

CERVICAL SWAB/URETHRAL SWAB:

• Dacron or Rayon swabs to be used
• Calcium alginate and Cotton swabs should not be used. Because Gonococci will die in
those swabs.
• Swabs should be transported in Stuart’s or Pike’s medium to maintain the viability of
bacteria

6. STOOL:
Indicated in:
Routinely in infants and children in case of diarrhea.
Adult: During endemic and sporadic diarrhea (bacterial, viral, parasitic).
Technique of collection:
1. Only one specimen per patient per day will be accepted.
2. Stool specimens should be collected in a clean, dry container
3. Transfer specimen from the collection container into a transport medium.
Do not overfill the transport vial.
4. Tighten the cap and shake well to mix.
5.Rectal swabs are substituted for stool culture specimens only if absolutely
necessary.
6.Do not refrigerate.
7. Send to the laboratory with proper labelling and filled-in request form.
Transport medium: Suspected cholera (V-R medium)
Suspected bacterial dysentery (Stuarts medium)
Processing:
• Macroscopic examination: Color, consistency, presence of blood.
• Microscopic Examination: Gram stain ( Pus cells, RBC )
Wet mount (Parasitic cysts / Oocysts, Trophozoite)
Hanging drop (Cholera bacilli)
Modified acid fast stain (cysts)
Commonly isolated/identified pathogens
Bacteria: Vibrio cholera, Salmonella, Shigella
Parasite: Isospora belli
Cysts & trophozoites of E. histolytica
Cysts of Giardia
Cryptosporidium parvum

7.CERIBROSPINAL FLUID
 Indications:
Pyogenic meningitis, Aseptic meningitis, Neonatal meningitis.
Technique of collection:
CSI is collected in sterile containers by lumbar puncture under aseptic condition
before institution of antibiotics in three different sterile containers.
Processing:
1st sample: Gram staining, India ink wet mount.
2nd sample: Biochemical analysis, Antigen testing by latex agglutination
3rd sample: Culture and sensitivity
Commonly isolated pathogens
▪ Staphylococci,
▪ Pneumococci,
▪ Meningococci,
▪ H.influenzae
▪ Cryptococcus neoformans.

8.BLOOD CULTURE:
Indicated in : Suspected Bacteremia, Typhoid fever
Technique of collection :
1.Wear sterile gloves.
2. Clean venipuncture site with 2% chlorhexidine using back and forth friction scrub for
30 seconds. Allow it to dry for 30 seconds. Do not retouch site
3. Hold pressure to site with gauze and apply adhesive bandage when bleeding stops
4. Remove venipuncture needle from syringe and fit with another sterile needle.
5. Remove tops of blood culture bottles and swab with chlorhexidine
6. Inject blood specimens into bottles, place specimen into the anaerobic Orange bottle,
then the aerobic Green bottle. If you only have enough blood for one bottle, place the
specimen in the aerobic Green bottle. Be careful, the bottles are pressurized and will
aspirate all of the blood from the syringe unless you control the flow with your thumb
and index finger.
7. Label the bottle.
8. Discard needle and syringe into a sharps container
9. Send specimen promptly to microbiology laboratory.
Processing :
Inoculate the specimen in blood culture bottle (Biphasic medium).
If growth appears, identify the isolate by appropriate biochemical tests.
Commonly isolated pathogens
• S. Typhi,
• Brucella,
• Staphylococci,
• Streptococci
• Pseudomonas aeruginosa.

SKIN SCRAPINGS:
 • skin scraping is a common procedure used to obtain the superficial dead layers of
the skin. This procedure is most often used to diagnose fungal skin infections, such as
athlete's foot or tinea infections of the body.
1. Clip 1cm2 of the area to scrape to remove hairs.
2.Squeeze the skin prior to scraping to push the mites out from the depths of the hair
follicles.
3.Scrape the skin in the direction of hair growth with a blade or spatula covered with
mineral oil until capillary bleeding is observed.
4.Put the scrapings in a sterile container, label it and send it to the laboratory with
properly filled in requisition form.

9. OPERATION THEATRE
Indicated in: To prevent nosocomial infections after any procedure in the Operation
theatre, to check for the presence of Clostridial spores
Technique of collection:
Swabs have to collected from specified areas and incubate in RCM broth,
incubate for one week to rule out the presence of Cl. tetani
Air sampling: Blood agar plates have to be exposed to OT environment for 1
hour and then incubate at 370C for overnight in the aerobic conditions. Check for
the MRSA, Pseudomonas and for fungi.