Pennsylvania Animal Diagnostic Laboratory System
PADLS User Guide - Contents Tests Mammalian Virology
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9.10 Mammalian Virology

Tests and assays are arranged by a weekly schedule, but designated emergency submissions are processed immediately upon arrival of the specimen for tests requested. The cell culture section maintains a variety of cell lines and SPF chicken embryonating eggs to provide isolation and identification of viruses from a variety of specimens from different species. Electron microscopy (negative staining or thin section) to identify viruses in tissue specimens is used where viruses are difficult to propagate or fail to replicate in cells, or to identify new pathogens by their morphology. The virology laboratory does not accept specimens from canine or feline species unless they are considered wildlife species or involved in a zoonotic problem.

Tissues for virus isolation should be collected as aseptically as possible. Viral and chlamydial transport media are available at no cost, upon request, from PADLS laboratories.

Note: Virus isolations may require several serial passages for amplification of replication and expression of viable and measurable virus in a specimen. Hence, certain virus isolations may require up to 28 days or longer. Identification of viral isolates is done using serologic or molecular techniques.

Procedure codes
AGID	agar gel immunodiffusion	FA	fluorescent antibody	PCR	polymerase chain reaction
CTM	chlamydia transport media	IHC	immunohistochemistry	VI	virus isolation
EM	electron microscopy	PAGE	polyacrylamide gel electrophoresis	VTM	viral transport media
LA	Latex Agglutination

Test/ Agent	Site	Procedure	Species	Tissue/ Specimen	Container	Shipping**	Turn- Around	Remarks
Adenovirus	PSU	VI	bovine, porcine	Nasal and ocular secretions*, feces, lung, brain, tonsil	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T.
Bluetongue virus; Epizootic hemorrhagic disease;	PSU	VI	deer, ovine	Vaginal secretions, serum from dam, fetal heart, heparinized blood*, spleen, bone marrow, lymph nodes, lung, semen	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T. Serotypes in USA: 2, 10, 11, 13, 17 PCR test done at NVSL
Border disease virus (hairy shaker): togavirus	PSU	VI FA	ovine	Brain, spleen, unclotted blood*, bone marrow, serum	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T.
Bovine enterovirus	PSU	VI EM	bovine	Feces*, oropharyngeal swab, intestine	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T.
Bovine mammillitis: herpesvirus	PSU	VI	bovine	Lesion scrapings, lesions, teat swab, fluid exudates from lesion*	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-21 days	Test is set up on T
Bovine respiratory syncytial virus	PSU	ELISA PCR	bovine	Fresh tissue collected less than 2 hours after death (trachea, lung*, nasal secretions, trachea wash)	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	ELISA: 5-6 days; PCR: 2-3 days	Virus is extremely labile to environmental conditions
Bovine viral diarrhea virus - earnotch	PSU PVL	IHC	bovine	1/4-1/2 inch full thickness ear punch or notch	Red top tube with formalin	Standard shipping	5-10 days	Samples in formalin should still be shipped within a week of collection
Bovine viral diarrhea virus - earnotch	PVL	ELISA	bovine	1/2 inch full thickness ear notch	Sterile container	Chilled/frozen overnight on ice	2 days	Samples can be submitted dry and should be shipped immediately after collection or frozen and shipped within 4 weeks Dry samples are preferred
Bovine viral diarrhea virus: mucosal disease	PSU	VI FA	bovine	Nasal secretions, oral lesions, lung, spleen, blood, mesenteric lymph nodes, intestinal mucosa, vaginal secretions, fetal tissues, whole blood	Leak-proof container w/VTM - Purple top EDTA vacutainer for whole blood.	Overnight shipment, on ice, do not freeze	VI: 7-14 days FA on tissue: 2-3 days	VI Test is set up on T.
	PSU	VI microplate test for persistently infected animals		VI microplate: 1 ml sterile serum	VI microplate: red top vacutainer	Overnight shipment on ice to PADLS-PSU	5-12 days	VI Microplate assay is set up on Monday. Samples must be received by 2:00 pm Friday
		ELISA antigen capture		ELISA: serum			5 days	ELISA is set up on Wednesday or by arrangement
		PCR¹		PCR: Tissues/serum			2 days	PCR¹ by arrangement at PSU
¹Call PSU (814.863.0838) for submission of more than 100 or for rush testing.
Chlamydia: psittacosis	PSU	FA VI	ovine, bovine, caprine	lung, liver, spleen, fetal tissues, placenta, cotyledon, nasal swab, feces/intestinal material	Leak-proof container with CTM	Overnight shipment, on ice, do not freeze	CI: 7-14 days	Test is set up on T
Contagious ecthyma (ORF): parapoxvirus	PSU	VI EM	caprine	Scabs, lesions on lip*	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days	Test is set up on T
Coronavirus-See neonatal diarrhea
Enteroviruses	PSU	VI EM	bovine, porcine	Vaginal secretions, serum from dam or sow, nasal swab, tonsil, brain (swine), feces (cattle and swine)	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days	Test is set up on T
Equine rhinopneumonitis: herpesvirus (EHV 1, 4)	PSU	VI FA	equine	Placenta-fetus, lung, nasal secretions, lymph nodes, thymus, brain	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days FA: 2-3 days	Test is set up on T
Equine rhinopneumonitis: herpesvirus (EHV 1, 4)	PSU PVL	PCR	equine	Unstained slide	slide mailer	Standard delivery	PCR: 5-10 days	Test is set up on T
Equine rhinovirus	PSU	VI	equine	Nasal secretions*	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T
Equine viral arteritis virus: togavirus	PSU	VI	equine	Whole blood, nasal and pharyngeal secretions, placenta, fetus, spleen, nostril, lymph nodes, conjunctival sac, semen	Leak-proof container with VTM Purple top vacutainer for whole blood	Overnight shipment, on ice, do not freeze	14-21 days	Test is set up on T
Hemagglutinating encephalomyelitis virus: coronavirus	PSU	VI HA	porcine	Brain*, spinal cord	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T
Infectious bovine rhinotracheitis virus	PSU	VI FA	bovine	Nasal and ocular secretions, lung, tracheal swab, tracheal segment, brain, vaginal secretions, serum, aborted fetus, liver, spleen, kidney	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days FA: 2-3 days	Test is set up on T
Influenza virus	PSU	VI HA FA	equine, porcine	Nasal and ocular secretions*, lung, tracheal swab	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days	Test is set up on T
Neonatal diarrhea - Coronaviruses	PSU	FA EM IHC	bovine	Feces 10 ml, small intestine*	Feces in leak-proof container Tissues for FA in VTM IHC in formalin	Overnight shipment, on ice, do not freeze	2-3 days	Do not use glass containers for feces as they may explode
Neonatal diarrhea - Coronaviruses	PVL	ELISA	bovine	Feces		Overnight shipment, on ice, do not freeze	2-3 days	ELISA is set up on T, Th
Neonatal diarrhea - Parvoviruses	PSU	VI EM	bovine, porcine	Feces, intestinal mucosa, regional lymph nodes, brain, heart	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days	Test is set up on T
Neonatal diarrhea - Rotavirus	PSU	EM	bovine, porcine	Feces, small intestine	Feces in leak-proof plastic container	Overnight shipment, on ice, do not freeze	2-3 days	Do not use glass containers for feces as they may explode
Neonatal diarrhea - Rotavirus	PVL	ELISA	bovine, porcine	Feces	Feces in leak-proof plastic container	Overnight shipment, on ice, do not freeze	2-3 days	Do not use glass containers for feces as they may explode
Papilloma virus	PSU	EM	bovine, equine	Lesion material, warts*	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	2-3 days
Parainfluenza virus: type 3 (PI₃)	PSU	VI FA PCR	bovine, ovine	Nasal and ocular secretions, lung, tracheal swab	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days FA: 2-3 days PCR: 2-3 days	Test is set up on T
Parvovirus See also neonatal diarrhea	PSU	VI FA	bovine, porcine	Vaginal secretions, serum from dam or sow, lung (mummified fetus)	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	7-14 days	Test is set up on T
Picornavirus enterovirus	PSU	VI EM	porcine	Feces, intestine, brain, tonsil, liver	Leak-proof container with VTM	Overnight on ice, do not freeze	7-14 days	Test is set up on T
Porcine Circovirus type 2 (PCV-2)	PSU	VI FA PCR	porcine	Lymph nodes, lung, spleen, tonsil	Leak-proof container with VTM	Overnight on ice, do not freeze	VI: 7-14 days PCR: 1-5 days
Porcine Circovirus type 2 (PCV-2)	PVL	IHC	porcine	Fixed tissue	Formalin	Standard	IHC: 10-14 days
Porcine inclusion body rhinitis: cytomegalovirus	PSU	VI	porcine	Turbinate, nasal mucosa	Leak-proof container with VTM	Overnight on ice, do not freeze	7-14 days	Test is set up on T
Porcine respiratory and reproductive syndrome: Arterivirus; Lelystad agent	PSU	FA VI PCR	porcine	Lung, tonsil, serum	Leak-proof container with VTM	Overnight on ice, do not freeze	FA: 2-3 days VI:14 - 21 days PCR: 5-6 days	Test is set up on T PCR is run on F
Poxviruses: cowpox, vaccinia, orthopox, parapox, pseudocowpox, ORF, bovine papular stomatitis	PSU	VI EM	bovine, porcine, rabbit	Lesion scrapings, lesions, vesicular fluids, crusts, liver, spleen	Leak-proof container with VTM	Overnight on ice, do not freeze	VI: 7-14 days EM: 2-3 days	Test is set up on T Debride lesions to obtain viable cells with virus
Pseudorabies: herpes virus	PSU	VI FA	porcine, ovine, bovine	Nasal secretions, tonsil, lung, brain (midbrain, pons, medulla), spinal cord (sheep and cattle), spleen (swine), vaginal secretion, serum	Leak-proof container with VTM	Overnight on ice, do not freeze	VI: 7-14 days FA: 2-3 days	Test is set up on T
Reovirus	PSU	VI	bovine, equine	Feces, intestinal mucosa, nasal and pharyngeal secretions	Leak-proof container with VTM	Overnight on ice, do not freeze	7-14 days	Test is set up on T
Rotavirus-see neonatal diarrhea
Transmissible gastroenteritis: coronavirus	PSU	EM FA	porcine	Feces, nasal secretions, jejunum, ileum	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze	VI: 7-14 days FA: 2-3 days	Test is set up on T
Unknown virus:	PSU	EM VI	any species	All tissues with lesions	Leak-proof container	Overnight shipment, on ice, do not freeze	3-45 days serial passages required	Test is set up when cell cultures are prepared
Vesicular stomatitis: rhabdovirus	PSU	VI	bovine, equine	Vesicular fluid, epithelial covering of lesions, whole blood, regional lymph nodes, tongue swab	Leak-proof container with VTM	Overnight shipment, on ice, do not freeze		Reportable virus
West Nile Virus	PVL	IHC PCR	all Species	Brain	Formalin fixed tissue		10-14 days

*preferred specimen
**If shipping to PSU, do not use the US Postal Service

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