Iowa Neonatology Handbook: Infection

Diagnosis of Infections Caused by Herpes Viruses and Chlamydia

Charles Grose, MD
Peer Review Status: Internally Peer Reviewed

Laboratory Hours and Location

Location: The University of Iowa Hospitals' Viral Diagnostic Laboratory is located in 265 Medical Research Center

Telephone Number: 356-4463

Hours: The University of Iowa Hospitals Viral Diagnostic Laboratory is open from 8:00 am to 4:30 pm Monday through Friday and from 8:00 to 11:00 am Saturday and Sundays. Specimens should be in the laboratory no later than 4:00 pm Monday through Friday for processing.

Receipt of Specimens After Hours: Every effort should be made to get specimens to the laboratory on the same day they are collected (by 4:00 pm). Overnight storage of specimens, especially urines and tissues, should be avoided; it is preferable to obtain the specimen early the next morning. When this is not feasible, specimens should be taken after 4:00 pm M-F and 0800 to 0800 S-S to 6248 RCP, Specimen Control. All specimens must be on ice.

Transport Media

Collection of the specimen into appropriate transport media is essential to ensure viability of the organisms to be cultured.

Transport Systems

Swabs: Virocult; Chlamydia Transwab
Available from Hospital Stores during regular working hours. Specimen Control during off-hours.

Tissue: Broth medium available in the Virology Lab, Specimen Control, 6248 RCP, Frozen Section Room.

Specimen Collection Guidelines For Clinical Virology Lab

1. Send all specimens to 265 MRC or Specimen Control 6248 RCP ON ICE.
2. Procedures Available:
   Isolation and Identification of Herpes simplex, cytomegalovirus, varicella-zoster, respiratory syncytial virus, Chlamydia trachomatis; rotavirus antigen detection; varicella-zoster immune status; and rubella immune status.
   1. Do not clean area to be cultured with alcohol.
   2. Viral
      1. Herpesviruses (CMV, HSV, VZV)
         1. Vesicle Fluid: Virocult swab of fluid obtained from disrupted vesicle or lesion.
         2. Spinal Fluid: Aseptic collection into sterile containers.
         3. Biopsy and autopsy tissue: Immerse tissue in broth transport media.
         4. Ulcerative lesions: Swabs (anal, genital, ocular). Use Virocult swab and scrape surface of lesion.
         5. Urine: Freshly passed AM urine is preferable. Collect clean voided midstream sample or suprapubic aspirate in sterile container. At least two or three specimens should be obtained to maximize recovery of CMV.
         6. Blood:
            • Rubella and Varicella serology: Submit at least 2 ml of clotted whole blood (red top tube) not necessary to hand carry or put on ice.
            • Buffy coat culture: Submit at least 1 ml of heparinized blood (green top tube).
      2. Respiratory Syncytial Virus: NP Washings, Tracheal aspirates, sputum; Throat swab as last resort.
      3. Human Immunodeficiency Virus (HIV)
         1. Blood
            HIV serology. Submit at least 2 ml of clotted whole blood (red top tube).
         2. Culture.
            Call State Hygienic Laboratory in Iowa City to obtain current instructions about methods for culturing HIV from blood or other specimens. Remember to obtain consent forms prior to submitting specimens.
   3. Chlamydia (trachomatis)
      1. For best results specimens should be aseptically collected with some vigor (swabbing or scraping) to insure an adequate number of epithelial cells in collection media.
         1. Swab: sterile Chlamydia Transwab * Conjunctiva: firmly stroke the everted lower eyelid with swab; place swab into transport media and transport to the lab on ice.
         2. Nasopharyngeal Wash: (Transwab acceptable)
            • Draw approximately 1-3 ml phosphate buffered saline (PBS) into an infant bulb syringe (or normal saline without preservatives).
            • Inject fluid into nostrils carefully.
            • Aspirate fluid from nostril into bulb syringe. Place contents into sterile tube.
         3. Urine: Not suitable for Chlamydia culture.
   4. Rotavirus Antigen Detection
      Liquid fecal specimen in sterile container. Swabs not acceptable.

Transport of Specimens

1. Specimen Transport:
   Carry all specimens ON ICE to:
   Virology Lab 265 MRC M-F 8 a.m. - 4:30 p.m.
   Specimen Control 6248 RCP all other times.
2. Storage of Specimens After Hours:
   Viral lability away from living cells necessitates that all viral specimens be transported on ice as rapidly as possible to the laboratory. When a specimen must be obtained after the laboratory is closed, specimens should be refrigerated at 4°C, if storage is less than or equal to 48 hrs. Otherwise specimens should be stored frozen at -70° C. Exceptions are urine specimens and tissue specimens which should not be frozen and ideally should be obtained the day they are to be cultured.

Clinical Disease Commonly	Associated with Virus	Clinical Specimens	Transport System
HSV - I, II	Mouth + Lips: gingivostomatitis recurrent herpes labialis	vesicle fluid or swab; throat swab	Virocult
	Eye: acute follicular conjunctivitis corneal ulceration stromal keratitis	vesicle fluid or swab;	Virocult
	CNS: encephalitis	Brain bx*; CSF; HSV PCR throat swab	Tissue Transport Media; Virocult
	aseptic meningitis	Buffy coat	Green top tube
	Skin: vesicle	vesicle fluid* or swab
	Generalized Infections: newborn	urine*; Buffy coat*; biopsy; vesicle swab*; throat swab*; bronchial washings; rectal swab*; conjunctivae	Heparinized (green top) tube Virocult
VZV	Chickenpox	vesicle fluid	Virocult
	Zoster	lesion swab or fluid (CSF)	Virocult
	Generalized Infections: newborn (pregnant mother with chickenpox)	serum	Red top tube
		throat swab, vesicle swab* or fluid, CSF*, urine, bronchial washings	Virocult
CMV	Generalized Infections:
	newborn	urine*, buffy coat;	Green top tube,
		saliva, CSF	Virocult
	CNS: infections	CSF, urine*
	Other: hepatitis, pneumonia, mononucleosis	urine*, buffy coat;	Green top tube,
		biopsy	Tissue Transport Media
Chlamydia Trachomatis	Eye: neonate and adult conjunctivitis	swab*	Chlamydia transport media
	Respiratory: neonate pneumonia	nasal wash*;	Chlamydia transport media
	? adult pneumonia	nasopharyngeal swab

* Recommended specimen for optimal growth in culture

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