Gram Stain


Abbrev Code:GRAM   
Order Code:LAB250Order Name:Gram Stain
Synonyms:GRAMST; Bacterial Smear
Methodology:Gram stain
CPT Codes: 87205 x1
Test Includes:Microscopic exam of Gram stained specimen. Preliminary review performed on all STAT requests and all CSF, subdural fluid, and subarachnoid fluid at UMMC/UMMCH West Bank, Southdale, Range, St. John's and Woodwinds prior to sending to the Infectious Diseases Diagnostic Laboratory (IDDL).
Turnaround Time:Performed 24 hours/day; results are reported in approximately 4 hours for routine testing, and 30-60 minutes for STAT testing.
Special Instructions:If ordering Aerobic Bacterial Culture or Respiratory Culture, do not order this test. Choose "with Gram stain" with Routine order. Indicate specimen site on request form.


Collection Instructions

Specimen:Fluid, tissue, aspirate, respiratory (sputum and bronchial) and swabbed anatomical sites.
Fluid, tissue and aspirate is always preferred over swab, when possible.
Automatically performed on: CSF, subdural fluid, subarachnoid fluid, sputum (expectorated), and vaginal. Other specimen types require a specific request.
Container:Plastic leakproof container, sterile, See Collection Instructions
Alternate Containers: CoPan ESwab with larger swab
Collection Instructions:
Specimen Type Instructions Volume Container Alternate Container
Abscess Prepare surface with povidone iodine solution; let dry.
Aspirate fluid/purulent material with syringe. Expel air bubbles from syringe; inject sample into sterile container
Deliver to laboratory immediately.		 Optimal: 5 mL
Minimum: 1 mL		 Sterile Container
 Eswab – for use only if fluid/material cannot be aspirated.
Aspirate
Cyst
Body Fluid:
Amniotic, Aqueous, Ascites, Gastric, Ocular, Pericardial, Pleural, Peritoneal, Synovial, Vitreous, Milk 
Semen Collect in sterile container
Washing
Bone Marrow Cultures should be taken after specimens for cell morphology studies, genetic studies, cell marker studies and other cell studies. Use a new syringe for each. Clean top of anticoagulant tube with alcohol wipe before injecting. Optimal: 4-8 mL
Minimum: 1.5 mL		 SPS
 Pediatric Blood Culture Bottle
Brushing Skin: Cleanse with 70% alcohol. Scrape off skin at active border of lesion; include some health skin. Place Brush in sterile container. N/A Brush in Sterile Container
 N/A
CSF Wear gloves, gown and mask. Prepare skin as for blood culture. Drape surrounding skin with sterile linen. Ideally, separate tubes should be collected for bacteriology, hematology cell counts and chemistries. Deliver to laboratory immediately. Optimal: 2mL
Minimum: 0.3 mL		 CSF Container
N/A
Bone:
Biopsy, Core Biopsy, Curetting, Fracture, Fragments, Resection, Exostosis, Osteophyte, Resection 		Place specimen in sterile leakproof container. Visible amount of specimen. Sterile Container
 Eswab – for use only if fluid/material cannot be aspirated.
Calculus/Stone
Foreign Body
Tissue:
Biopsy, Fine needle aspiration
Placenta Place patient in lithotomy position, insert speculum, and visualize cervical os.
Remove mucous plug, wipe cervical os with povidone iodine solution. Wipe off povidone iodine solution.
Aspirate material through the sleeve of a rush or curette sampling implement to avoid normal flora contamination.
Place the aspirate in a sterile tube.
Do not collect on swabs to prevent contamination with normal cervical/vaginal flora and to ensure adequate material is collected. Specimen can be swabbed after removal if shared with Surgical Pathology.
Anaerobic container can be used for aerobic and anaerobic culture.		 Visible amount of specimen. Sterile Container Eswab – for use only if shared with surgical pathology.
Swab Clean Surface with sterile sponge and sterile water.
If purulent material is available to be Aspirated collect in Sterile container for improved sensitivity. 		Eswab- N/A
Sterile Container:
Optimal: 5 mL
Minimum: 0.5 mL 		Sterile Container
 Eswab – for use only if fluid/material cannot be aspirated.

Culturette Swab: FRMC Only
Skin
Wound
Specimen Type Instructions Volume Container Alternate Container
Aspirate (Tracheal)  
Collect in sterile leakproof container.
 		 Optimal:
3 mL
 
Minimum:
0.5mL		 Sterile Container
 N/A
Bronchial Alveolar Lavage
Washing (Bronchial)
 
Brushing (Bronchial) Bronchial Brush: Using standard bronchoscopy technique, identify the lesion in question and obtain a brushing sample of the lesion. Upon withdrawing the brush, agitate the brush vigorously in a 5-10 mL vial of sterile saline. Transport to the laboratory immediately, specimen should arrive within 3 hours.
 
Sputum
 		 Expectorated sputum is not recommended for routine bacterial/fungal culture because of unavoidable contamination with upper respiratory flora. If possible, have the patient rinse mouth and gargle with water first. Ask the patient to cough deeply and expectorate coughed-up material into a sterile container. In seriously ill or debilitated patients, a transtracheal aspirate may be necessary.
Causes for Rejection:  More than 10 epithelial cells per low power field, appearances of saliva.
Sinus Contents or Stripping (Sinus) Clean Surface with sterile sponge and sterile water.
If purulent material is available to be Aspirated collect in Sterile Container for improved sensitivity.
 		 Optimal:
3 mL
 
Minimum:
0.5mL
 		 Sterile Container
 Eswab
Throat Swab Collect one swab; collect second swab if also collecting rapid strep screen. Place each swab in culture container to ensure proper preservation of specimen. Collection technique for throat: Depress tongue and rub swab vigorously over each tonsillar area and posterior pharynx. Any exudate should be touched, and care should be taken to avoid tongue and uvula. N/A Eswab White
Eswab Blue
Ear Swab Place swab in culture container to ensure proper preservation of specimen. Specimen can be divided for aerobic culture or fungus culture and stain if ordered and there is sufficient volume.
  • INTERNAL: Cleanse external canal with mild antiseptic. Insert sterile funnel and swab from ear drum or beyond.
  • EXTERNAL: Cleanse external canal with mild antiseptic. Swab or scrape from active margin, preferably including fresh secretion from deeper areas.
 N/A Eswab Blue

 		 Eswab White

 
Causes for Rejection:Not an adequately sensitive test on direct blood films; not useful on throat or fecal specimens except to screen for overgrowth of yeast. Gram stain may not be possible on hard bone specimens or foreign bodies (i.e., catheter tips) without the presence of soft tissue or purulent material. Testing on these specimens may be canceled at laboratory's discretion.


Processing and Shipping

Specimen Processing:Store at room temperature.
Shipping Instructions:Ship slides at room temperature in protective slide holder. If a culture is also requested, laboratory will use the original specimen to prepare a smear and a separate slide is not required.
Test Performed at or Referral Lab Infectious Diseases Diagnostic Laboratory   (Infectious Diseases Diagnostic Lab-Micro (UMMC East Bank), Microbiology (Grand Itasca))


Interpretive

Reference Range:No organisms seen.
Use:Provides preliminary information regarding presence or absence of bacteria, yeast, neutrophils, epithelial cells, pathogenic organisms, anaerobic organisms, Sensitivity of culture is significantly higher as compared to gram stain alone. Culture also allows antibacterial susceptibility testing of pathogenic organisms. It is recommended to order appropriate culture in conjunction with gram stain. Diagnosis of bacterial vaginosis does not required a culture. May be helpful for diagnosis of moniliasis (thrush).


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