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Virology Index
  Specimen Collection Instructions
Table 1: Virology Specimen Collection for Viral Cultures, Direct Exam (DFA) and Rapid Antigen Testing
Specimen Submission
Table 2: Viruses Associated With Disease Categories
Nasopharyngeal Specimen Collection
Conjunctival Specimen Collection for Detection of Viruses and Chlamydia
Collection of Lesion Samples for HSV and VZV Viral Detection

Virology

Specimen Collection Instructions

Table 1: Virology Specimen Collection for Viral Cultures, Direct Exam (DFA) and Rapid Antigen Testing (Refer to Test Directory for Molecular Testing Requirements)

Specimen Source/Type

Collection Container

Representative Viruses

Comments

Blood

10 mL in Yellow-top  tube (ACD) Acid Citrate Dextrose, Solution A

CMV, HSV, VZY, Adenovirus, Enterovirus

Specify suspected virus on requisition. If no virus is specified, only CMV PCR testing will be performed. Consider Molecular testing for specific viruses.

Bone marrow

2 mL in Yellow-top tube (ACD)  Acid Citrate Dextrose) or Blue-top tube (Sodium Citrate)

CMV

 

Eye, conjunctiva/cornea

Swab inside of lower, then upper lid. Collect infected epithelial cells not pus. Place 1-2 swabs, fluid or tissue into VTM tube. 

Adenovirus, HSV, Chlamydia, Enterovirus, CMV, VZV

Request both viral culture and DFA for optimal detection of Adenovirus, HSV and VZV.  If both tests are requested, must collect 2 swabs. Both swabs can be placed into 1 VTM tube.
Chlamydia culture is a separate test request.

Fluids

2-5 mL in sterile screw-cap tube

 Varies with source

 

CSF

1-5 mL in sterile screw cap tube

Enterovirus, HSV, Mumps, CMV, VZV, Measles, Influenza

Mumps or Measles culture must be specified on requisition. Consider Molecular testing for specific viruses.

Pericardial fluid

1-5 mL in sterile screw cap tube

Enterovirus, CMV

Consider Molecular testing for specific viruses
Request PCR for enterovirus, HSV, CMV, VZV, etc.

Genital swabs (cervix, vulva, urethra, penis)

Place 1-2 swabs into VTM tube. Collect adequate cells from infected area. Collect both fluid and cells from base of lesion.

HSV, CMV (Chlamydia)

Consider Molecular testing for Chlamydia.

Lesions, derma/mucosal vesicles

Place 1-2 swabs into VTM tube.  Collect both fluid and cells from base of lesion.

HSV, VZV,  Enterovirus

Request both culture and DFA for optimal detection. If both tests are requested, must collect 2 swabs. Both swabs can be placed into 1 VTM tube.

Respiratory
- NP/throat swabs
- Lung tissue
- BAL, other fluids

Place 1-3 swabs aspirates or tissue into VTM tube.

Fluids including BAL should be placed into sterile screw-cap tube

Adenovirus, CMV, HSV, RSV, Enterovirus, Influenza, Mumps, Parainfluenza, Rhinovirus, Rubeola, Rubella, VZV,  human Metapneumonovirus, (Chlamydia in neonates)

Rapid Antigen Testing (STAT) requires 1 swab in dedicated VTM tube.
Culture and Direct Slide (DFA) requests require 2 swabs, which can be combined into 1 VTM tube.
Consider molecular testing for human Metapneumonovirus, CMV or Chlamydia. Measles, mumps, and rubella must be specified on Requisition.

Stool

Place walnut size amount into a Screw-cap container

Adenovirus, Enterovirus, HSV, VZV

Adenovirus types 40/41 and Rotavirus are not detected by culture and must be requested individually.

Rectal

Place swabs into VTM tube.

Enterovirus, HSV, VZV

 

Tissues/Biopsy

Place tissue (3 mm or more) into VTM tube.

HSV, CMV, VZV
Varies with source

Consider Molecular testing for specific viruses.

Urine

Place into Sterile screw- cap container

CMV, Adenovirus, Enterovirus, HSV, Mumps

Mumps by special request only.

VTM - viral transport medium. Once specimens have been collected, keep refrigerated and transport promptly.

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Specimen Submission

Request Forms
Use individual requisition forms available from the laboratory.  All test requests require a physician's written order to process a specimen. Follow the collection instructions for each type of specimen.

Patient Identification
All patients from whom clinical specimens are obtained must be positively identified, utilizing at least two unique identifiers prior to specimen collection.  Positive identification is the responsibility of the person collecting the sample.

Required Information
All specimens must be labeled.

Specimen Labeling: The following information must be legibly recorded on a label affixed in an irreversible fashion to the specimen container:

  • Patient’s full name (not a nickname)
  • Medical Record Number or other unique identifier (ID)
  • Date and, if appropriate, time when specimen was obtained
  • Specimen source
  • Signature/ initials of collector
  • The label should be affixed directly to the specimen container and not the bag

Bar-coded pre-printed labels with accession numbers generated by an information system may be used.

  • Place the labeled specimen in the provided leak proof sealed plastic biohazard bag
  • Place the matching requisition in the outside pouch of the bag
Transport specimens promptly: See specific test for temperature requirements
The date and signature/ initials of the collector must be recorded after the specimen has been collected and after verifying that the patient name and ID on the label agrees with that on the test requisition.  This is the single most important factor in preventing errors in patient specimen identification.

Use of a request form wrapped around the container is not acceptable as a specimen label.

Specimens will not be accepted if the information on the specimen label does not match the information on the accompanying requisition.

Required Information on the Requisition Form

On all requests forms, the following information is required

  • Patient's name & address
  • Patient’s gender
  • Date of birth
  • The last six digits of the patient’s social security number or other unique identifier (ID#)
  • Date and if appropriate, time of collection
  • Test requested
  • Type or source of the specimen
  • Requesting physician/ or Client Number
  • Clinical information if requested
  • All applicable medical necessity codes (ICD-9)
  • Complete billing and insurance information

Providing additional relevant information may be important in alerting the laboratory of the need for special handling or specimen work-up.

Tests sent to reference laboratories must have patient history information.  The need for such information is indicated on the test request form.

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Table 2: Viruses Associated With Disease Categories

Disease Category

Common Virus

Less Common

CNS

 

 

    Aseptic meningitis

Coxsackie, Echovirus, Mumps, HHV-6

Other enteroviruses, HSV-2, HIV, LCM, VZV

    Paralysis

Polio

Coxsackie, Enterovirus 71, other Enteroviruses

    Encephalitis

Arboviruses, HSV, Enteroviruses, HIV, CMV, HHV-6, Mumps

Herpes B, CMV, PML viruses (JCV), Rabies, VZV

Genitourinary

 

 

    Lesions

Herpes, Molluscum, HPV

CMV, VZV

    GU tract

CMV

Mumps, HSV

    Cystitis, Glomerulonephritis

Adenovirus, BKV

 

Gastrointestinal

Adenovirus, CMV, norwalk, rotavirus

Enteroviruses

Hepatitis

Hepatitis A, B, C, D, EBV, CMV

Hepatitis E, G, TTV, HSV, Togavirus (Yellow Fever),  Arenavirus

Neonatal disease

 

 

    TORCH

Rubella, CMV, HSV, Echovirus, Parvovirus, Hepatitis B

Adenovirus, VZV

    NB respiratory

Influenza, HSV, RSV

Adenovirus, Measles, Parainfluenza, Enteroviruses

Ocular

 

 

    Conjunctivitis

Adenovirus, enteroviruses, HSV, VZV, (Chlamydia)

Dengue, Newcastles

    Corneal Lesion

HSV

Respiratory

 

 

    Upper respiratory

 

 

        Colds, Pharyngitis

Adenovirus, Coxsackie, EBV, Echovirus, Influenza A & B, Respiratory Syncytial (RSV), Herpes, Parainfluenza, Rhinovirus

Coronavirus, Influenza C, Parainfluenza

        Croup, Bronchiolitis

RSV, Parainfluenza

Adeno, Influenza, Measles

    Lower Respiratory

 

 

        pneumonia, adults

Influenza

Adenovirus, measles, newcastles; immunocompromised: CMV, HSV, VZV

        pneumonia, children

RSV, influenza, parainfluenza

Adeno, measles, VZV

Skin

 

 

    hemorrhagic

Arboviruses

Ebola

    lesions, local

HSV, VZV, HPV, molluscum

Poxviruses

    maculopapular

Coxsackie, echovirus, parvovirus, rubella, rubeola (measles)

Adenoviruses, CMV, EBV

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Nasopharyngeal Specimen Collection

Specimen Collection for Detection of Respiratory Viruses (including Adenovirus, CMV, Enterovirus, Influenza, Parainfluenza, RSV, and Rhinovirus).  Other appropriate specimens include throat/pharyngeal swabs, tracheal aspirates, and bronchial samples.

Place fluids in sterile leakproof containers and transport to the laboratory.

Nasopharyngeal Swab Method

Materials:

  • Nasopharyngeal swab with synthetic fiber tip
  • 1-2 mL viral transport medium (VTM)
  • Specimen container

Do not use calcium alginate swabs. Dacron swabs are provided with viral transport medium.
nasopharyngeal specimen collection

  • Insert swab into one nostril.
  • Press swab tip on the mucosal surface of the midinferior portion of the inferior turbinate (see sketch), and rub the swab tip several times across the mucosal surface to loosen and collect cellular material.
  • Withdraw swab; insert into container with VTM.

For best sample quality, repeating procedure for the second nostril will deliver optimal combined sample.  Collect two to three swabs of NP and/or pharynx.  Place swabs into viral transport medium. Tighten caps(s) securely. Samples must be kept cold and received within 24 hours of collection. Submit specimens promptly to the laboratory.  Refrigerate
For collection kits. call Customer Services at 1- (877) 717-3733.

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Conjunctival Specimen Collection for Detection of Viruses and Chlamydia (Adenovirus, Chlamydia trachomatis, Enterovirus, and Herpesvirus)

  • Gently remove pus or discharge. Use for Gram stain or microbial cultures only.
  • Swab inside of lower, then upper lid as pictured. Collect infected epithelial cells (not pus) on swabs for viral and chlamydial detection.
  • Use dacron swabs supplied with viral transport medium (VTM). Do not use calcium alginate or wood swabs as these are inhibitory to viruses and Chlamydia.
  • Collect multiple swabs for both eyes (if affected) and place in viral transport medium (VTM). Collection of throat or nasopharyngeal swabs may increase detection of viruses.
  • Note: For detection of HSV from corneal ulcers, collect cells from affected area of cornea also and submit in viral transport medium (VTM).
  • Submit specimens promptly to the laboratory.
  • Eye specimens can be tested for the following agents:
    • Adenoviruses
    • Chlamydia trachomatis
    • Enteroviruses (Coxsackie, echovirus, poliovirus)
    • Herpes simplex virus

Collect multiple swabs if both culture and direct viral exams are desired. Two or three swabs can be submitted in each vial of viral transport medium (VTM).

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Collection of Lesion Samples for HSV and VZV Viral Detection

To increase the chance of obtaining positive results, it is necessary to collect the types of cells that the HSV typically infect.

  • In figure 1, normal epithelium can be seen to consist of four distinct zones.
  • In figure 2, these same zones are seen in herpetic lesions.

The non-superficial cells, intermediate, para-basal, and particularly the base cells - are the ones that can become infected. To obtain these cells it is critical that the base of the lesion be thoroughly scraped. If the laboratory sees only superficial cells, erythrocytes, or polymorphonuclear leukocytes, the sample is inappropriate for this test.


Figure 1



Figure 2

Collection Procedure

  • To expose the base of lesion:
    • Vesicles: Use a sterile needle to lift up the cap of the vesicle. Note that while vesicular fluid is not an adequate sample for the direct test, it is ideal for isolation. If vesicular fluid is present, aspirate with a sterile needle and syringe and inject into an appropriate transport medium (VTM).
    • Ulcers: Use a sterile swab to remove any unwanted pus without disturbing the base of the lesion; discard the swab after use.
    • Crusts: Use a sterile needle to expose the base of the lesion.
  • Moisten a large or small dacron swab in sterile water and vigorously swab the entire base of the lesion.  Vigorous scraping of the lesion base is essential and this can be expected to cause the patient momentary pain.
  • Collect two to three lesion swabs and place in viral transport medium. Tighten cap securely. Transport promptly to the laboratory.

Samples must be kept cold and received within 24 hours of collection.
Submit specimens promptly to the Laboratory (24 hours/7 days).

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