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Chapter 10.4 : Specimen Collection and Processing

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Abstract:

Successful viral culture requires careful attention to the selection, collection, transport, and assessment of specimens. The information contained in this procedure is essential not only to those laboratories performing viral cultures on site but also to those outsourcing specimens. The laboratory must provide written guidelines to collection sites (nursing station, clinic, physician's office, and emergency room) detailing cultures that are available and instructions for submitting specimens. Instructions should include laboratory hours of operation and contact person(s); instructions for specimen collection, labeling, storage, and transport; source and storage conditions for transport media and containers; information required for adequate testing; turnaround time; reporting procedures and values; and testing limitations.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Figures

Image of Figure 10.4-1
Figure 10.4-1

Summary of collection, transport, and evaluation of specimens for viral culture. NPA, nasopharyngeal aspirate; BAL, bronchoalveolar lavage fluid.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Image of Figure 10.4-2
Figure 10.4-2

Preparation of fluid specimens. soln, solution; NPA, nasopharyngeal aspirate; TA, tracheal aspirate; BAL, bronchoalveolar lavage fluid.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Image of Figure 10.4-3
Figure 10.4-3

Preparation of swabs, scrapings, and vesicle aspirates collected in VTM.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Image of Figure 10.4-4
Figure 10.4-4

Preparation of tissue samples.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Image of Figure 10.4-5
Figure 10.4-5

Preparation of stool samples. tsp, teaspoons.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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Image of Figure 10.4-6
Figure 10.4-6

Preparation of leukocytes from anticoagulated blood by the ammonium chloride method. For details, see reference 10. PBS, phosphate-buffered saline.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
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References

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1. Clarke, L. M.,, B. J. Daidone,, R. Inghida,, M. Kirwin,, and M. F. Sierra. 1992. Differential recovery of cytomegalovirus from cellular and supernatant components of bronchoalveolar lavage specimens. J. Clin. Pathol. 97:313317.
2. Crane, L. R.,, P. A. Gutterman,, T. Chapel,, and A. M. Lerner. 1980. Inoculation of swab materials with herpes simplex virus. J. Infect. Dis. 131:531.
3. Eggleton, P.,, R. Gargan,, and D. Fisher. 1989. Rapid method for the isolation of neutrophils in high yield without the use of dextran or density gradient polymers. J. Immunol. Methods 121:105113.
4. English, D.,, and B. R. Anderson. 1974. Single-step separation of red blood cells, granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J. Immunol. Methods 5:249259.
5. Ferrante, A.,, and Y. H. Thong. 1980. Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leukocytes from human peripheral blood by the Hypaque-Ficoll method. J. Immunol. Methods 36:109117.
6. Howell, C. L.,, and M. J. Miller. 1983. Effect of sucrose phosphate and sorbitol on infectivity of enveloped viruses during storage. J. Clin. Microbiol. 18:658662.
7. Howell, C. L.,, M. J. Miller,, and D. A. Bruckner. 1986. Elimination of toxicity and enhanced cytomegalovirus detection in cell cultures inoculated with semen from patients with acquired immunodeficiency syndrome. J. Clin. Microbiol. 24:657660.
8. Howell, C. L.,, M. J. Miller,, and W. J. Martin. 1979. Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/ Macrodex methods. J. Clin. Microbiol. 10:533537.
9. Johnson, F. B. 1990. Transport of viral specimens. Clin. Microbiol. Rev. 3:120131.
10. Menegus, M. A.,, C. M. Mayer,, C. F. Mellen,, and J. R. Zeller. 1994. A simple and rapid method for the preparation of white blood cells (WBC) suitable for CMV culture. Abstr. Tenth Annual Clinical Virology Symposium, Clearwater, Fla..
11.U.S. Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. 1999. Biosafety in Microbiological and Biomedical Laboratories. Publication no. (NIH) 93-8395, 4th ed. U.S. Government Printing Office, Washington, D.C..
12. Warford, A. L.,, R. A. Levy,, K. A. Rekrut,, and E. Steinberg. 1986. Herpes simplex virus testing of an obstetric population with an antigen enzyme-linked immunosorbent assay. Am. J. Obstet. Gynecol. 154:2128.

Tables

Generic image for table
Table 10.4-1a

Specimen collection for viral cultures

Certain agents that may be present in clinical specimens (e.g., avian influenza virus, SARS-CoV, variola virus) require special handling and/or a biosafety level that exceeds that of many clinical laboratories. The laboratory must provide information to individuals submitting specimens regarding these issues and emphasize the importance of providing clinical history information, including pertinent travel, animal contact, and/or exposure history. Information is available at http://www.cdc.gov and http://www.asm.org, including at the following links: http://www.asm.org/Policy/Index.asp?bid_6342, http://www.cdc.gov/ncidod/SARS/lab.htm, http://www.cdc.gov/flu/avian/professional/han020302.htm, and http://www.bt.cdc.gov/agent/smallpox/diagnosis/rashtestingprotocol.asp. Abbreviations: tsp, teaspoons; HHV-6, human herpesvirus type 6; HIV-1, HIV type 1; HSV, herpes simplex virus; LCMV, lymphocytic choriomeningitis virus; VZV, varicella-zoster virus.

Use sterile screw-cap containers for fluid (other than blood) and bulk samples. For collection of swab samples, Dacron-tipped plastic- or wire-shafted swabs are recommended; do not use cotton or wooden-shafted swabs.

Includes group A and group B coxsackieviruses, echoviruses, enterovirus types 68 to 71, and polioviruses.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
Generic image for table
Table 10.4-1b

Specimen collection for viral cultures

Certain agents that may be present in clinical specimens (e.g., avian influenza virus, SARS-CoV, variola virus) require special handling and/or a biosafety level that exceeds that of many clinical laboratories. The laboratory must provide information to individuals submitting specimens regarding these issues and emphasize the importance of providing clinical history information, including pertinent travel, animal contact, and/or exposure history. Information is available at http://www.cdc.gov and http://www.asm.org, including at the following links: http://www.asm.org/Policy/Index.asp?bid_6342, http://www.cdc.gov/ncidod/SARS/lab.htm, http://www.cdc.gov/flu/avian/professional/han020302.htm, and http://www.bt.cdc.gov/agent/smallpox/diagnosis/rashtestingprotocol.asp. Abbreviations: tsp, teaspoons; HHV-6, human herpesvirus type 6; HIV-1, HIV type 1; HSV, herpes simplex virus; LCMV, lymphocytic choriomeningitis virus; VZV, varicella-zoster virus.

Use sterile screw-cap containers for fluid (other than blood) and bulk samples. For collection of swab samples, Dacron-tipped plastic- or wire-shafted swabs are recommended; do not use cotton or wooden-shafted swabs.

Includes group A and group B coxsackieviruses, echoviruses, enterovirus types 68 to 71, and polioviruses.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4
Generic image for table
Table 10.4-1c

Specimen collection for viral cultures

Certain agents that may be present in clinical specimens (e.g., avian influenza virus, SARS-CoV, variola virus) require special handling and/or a biosafety level that exceeds that of many clinical laboratories. The laboratory must provide information to individuals submitting specimens regarding these issues and emphasize the importance of providing clinical history information, including pertinent travel, animal contact, and/or exposure history. Information is available at http://www.cdc.gov and http://www.asm.org, including at the following links: http://www.asm.org/Policy/Index.asp?bid_6342, http://www.cdc.gov/ncidod/SARS/lab.htm, http://www.cdc.gov/flu/avian/professional/han020302.htm, and http://www.bt.cdc.gov/agent/smallpox/diagnosis/rashtestingprotocol.asp. Abbreviations: tsp, teaspoons; HHV-6, human herpesvirus type 6; HIV-1, HIV type 1; HSV, herpes simplex virus; LCMV, lymphocytic choriomeningitis virus; VZV, varicella-zoster virus.

Use sterile screw-cap containers for fluid (other than blood) and bulk samples. For collection of swab samples, Dacron-tipped plastic- or wire-shafted swabs are recommended; do not use cotton or wooden-shafted swabs.

Includes group A and group B coxsackieviruses, echoviruses, enterovirus types 68 to 71, and polioviruses.

Citation: Garcia L. 2010. Specimen Collection and Processing, p 40-50. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.4

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