132 Section IV ■ Miscellaneous Sampling

(3) For chlamydial PCR: Place specimen in transport medium appropriate for the assay used. An

example is M4 medium for the transport of

viruses and Chlamydia (Remel, Lenexa, Kansas).

2. Media for culture

 Specimens should be plated onto culture medium at

the bedside. Each laboratory will have specific media available for a particular type of organism. The following list is a

suggestion of classic media used for each type of organism.

a. Bacterial culture media

(1) Trypticase soy broth

(2) Blood agar plate

(3) Chocolate agar plate for Haemophilus influenzae,

Neisseria gonorrhea

(4) Thayer-Martin medium if gonorrhea suspected

b. Virus-holding medium (i.e., M4 medium for the

transport of viruses and Chlamydia) (Remel,

Lenexa, Kansas)

c. Chlamydia culture transport medium (i.e., M4

medium for the transport of viruses and Chlamydia)

(Remel, Lenexa, Kansas)

d. Sabouraud’s agar if fungal conjunctivitis suspected

F. Technique

1. Method for staining the cornea for epithelial defect

a. Instill a very small amount of fluorescein in the lower

conjunctival fornix by lightly touching the tear film

with a fluorescein strip. Flooding the eye with fluorescein may obscure a small corneal epithelial defect.

b. Evaluate the cornea for staining with a Wood lamp

or other blue light source.

c. If a corneal epithelial defect is present, the cornea

may be infected and an ophthalmologist should be

consulted.

d. Herpes virus may present in the neonate as a geographic-shaped epithelial defect rather than a dendrite.

2. Method for everting eyelids

a. Upper lid (Fig. 24.1)

(1) Grasp lashes and border of lid between thumb

and index finger of nondominant hand.

(2) Draw lid downward and away from eyeball.

(3) Indent upper lid, with handle of cotton-tipped

applicator held in dominant hand and pull lid

back and upward over applicator.

(4) Remove applicator and hold lid in place with

nondominant hand by gently pressing border of

lid against superior orbital margin.

b. Lower lid (Fig. 24.2)

(1) Place index finger of nondominant hand on

margin of lower lid.

(2) Pull downward.

3. Method for obtaining cultures

 Obtain cultures prior to conjunctival scraping. Take

separate cultures from each eye with a separate sterile

swab for each type of medium desired. Culture and

label each eye separately, even if only one eye is symptomatic. The uninfected eye can serve as a control for

indigenous flora (9).

a. Moisten calcium alginate swabs with trypticase soy

broth or other liquid culture medium.

b. Evert eyelid.

c. Apply swab to bulbar and palpebral conjunctiva of

upper and lower fornices of eye.

d. Apply swab directly to culture medium plates at the

bedside with a single row of C-shaped inoculation

streaks. Monitoring the growth of organisms along

the shape of the inoculation streaks may help the

laboratory in the diagnosis of the cultured pathogen.

e. Use a separate sterile swab for each culture plate or

culture vial.

f. Label cultures meticulously with eye cultured (right

or left) and part of eye cultured (conjunctiva, lid margin, etc.).

g. Incubate cultures immediately.

4. Method for obtaining conjunctival scrapings for smear

and nonculture Chlamydia tests

a. Evert eyelid as described above.

b. Instill topical anesthetic into conjunctival fornix, if

desired.

Fig. 24.1. Everting the upper eyelid.

Fig. 24.2. Using Kimura platinum spatula to take scraping from

lower eyelid.