IMPORTANCE OF SURGICAL SPECIMEN HANDLING IN INFECTION CONTROL

BY

DR AMJAD ALI KHAN

PRESENTATION FORMAT

Specimen collection. Specimen fixation. Specimen transportation. Handling the spills. Special situations.

SPECIMEN COLLECTION:
Wash hands prior to collection of specimen. Clean hands by applying sanitizer. Wear gloves. Put the specimen in the container with the help of forceps. Add formalin to the specimen in the container in the ratio of 15 to 20 :1. Tighten the cap. Put the container in the biohazard bag that is then zipper locked. Put the request form in the pouch attached.

SPECIMEN CONTAINERS
1. Containers should be rigid, impermeable, unbreakable and non-reactive to fixative. 2. The correct size of container must be used to hold and protect specimens. 3. Containers should have a secure, tight-fitting cover/lid to prevent the fixative solution from escaping. 4. IV solutions bottles should not be used for specimen collection and transportation.

Fixation (formalin based fixatives):

The aim of fixation is to stop decay, putrefaction and autolysis of tissues. It works by cross linking of proteins and aldehydes. The formalin added to the tissue should be in the ratio of 15 to 20:1. The speed of penetration by formalin is 0.5 mm per hour at room temperature. Fixation is hampered by fat, pus, necrotic tissue, food matter and blood. Almost all infectious agents are killed by formalin fixation except: Prions. Micro-organisms at the center of a large vascular or necrotic tissue, like spleen, placenta etc.

SPECIMEN TRANSPORTATION
The sealed specimen container should be transported to the laboratory as soon as possible. Delivery at the laboratory should be documented. Precaution should be taken to avoid any spills.

SPILLS IN THE LABORATORY:

Hold your breath, leave the room immediately, and close the door. Warn others not to enter the contaminated area. Remove contaminated garments and put them in a container for autoclaving. Thoroughly wash hands and face and any other exposed areas of the body. Wait 30 minutes to allow aerosols created by the spill to settle if in a laboratory where the airflow is negative. Put on a long-sleeve gown, mask, and rubber gloves before re-entering the room. Pour a decontaminant solution like concentrated bleach around the spill and allow to flow into the spill. Paper towels soaked with the decontaminant may be used to cover the area. Let stand for 20 minutes to allow an adequate contact time. Transfer all contaminated materials (paper towels, glass, liquid, gloves, etc.) into a deep autoclave pan. Cover the pan with aluminum foil.

Transmissible Spongiform Encephalopathies (TSEs)

Kuru Gertsmann-Straussler-Scheinker (GSS) Fatal Familial Insomnia (FFI) Creutzfeldt-Jakob Disease (CJD) Variant CJD (vCJD)

Handling cases of suspected Prion disease (history of subacute or chronic dementia):

Formalin fixation alone is ineffective in reducing the infectivity of prions. Gloves and protective eye shields are to be worn at all stages of handling suspected tissues. To inactivate prions: Place brain biopsies in 10% neutral buffered formalin for 48 hours. Place the fixed tissue sections 4-5 mm. in 50-100 ml of formic acid for one hour. Re-immerse the tissues in 10% buffered neutral formalin for 48 hrs, followed by tissue processing. Place the instruments in 2N NaOH sol. and for 01 hour at 134o C at pressure of at least 30lbs/m2

Tissue for Frozen Section:

1. Handle all tissues with gloves. 2. Discarded later in the orange bio-hazard bag. 3. Section the tissue as usual in the cryostat. Aerosols used for fast freezing are contraindicated and are not used. 4. Clean up and sterilize instruments and contaminated surfaces as follows: a. Cryostat - Daily wipe all surfaces with a towel or sponge which has been soaked in absolute ethanol. b. Cryostat Knife Holder - monthly soak in pan of ethanol for 15 min. c. Dissection instruments - Daily soak in 1/10 dilute bleach for 15 min. then wash well in tap water. d. Bench surfaces- Daily use standard bench disinfection technique.

RISK DURING GROSSING:

Properly fixed specimens carry low risk of infection. Poorly fixed and unfixed specimens carry high risk of infection. Tissue like placenta, spleen, lung and liver are not properly fixed until thin slices are put in formalin for a longer period of time. Splashing of liquids or specimens can produce aerosols. Cutting of bone by saw can also produce aerosol. Clean the cutting board by 1:10 liquid bleach. Wash the used instruments and soak them for 01 hour in liquid bleach.

CONTAMINATED SPECIMENS:
Placenta and spleen: Large vascular structure, contain large amount of blood and it takes much longer time for fixation. Infecting organisms in them might persist for a long time. Lung: Due to its air content it floats on the surface and does not fix well. Infecting organisms like Mycobacterium might be viable for long time. Diabetic gangrenous amputation specimens: May contain mixed population of different aerobic and anaerobic bacteria. The pus and necrotic material slows down the rate of fixation. Brain and spinal cord specimens: Patients with suspected sub-acute or chronic dementia may contain prions that cause slow virus diseases and are not killed by formalin.

ALL SURGICAL SPECIMENS SHOULD BE TREATED AS POTENTIALLY INFECTIOUS

THANK YOU