Viral Specimen Selection and Collection Overview
Viral Specimen Selection and Collection:
Specimen selection depends on the specific disease syndrome, viral
etiologies suspected and time of year.
1.Throat,
nasopharyngeal swab or aspirate: Throat swabs are acceptable for
recovering enteroviruses, adenoviruses and HSV, whereas nasopharyngeal
swab or aspirate specimens are preferred for the detection of influenza and
parainfluenza viruses. Throat specimens are collected by rubbing inflamed,
vesiculated, or purulent areas of the posterior pharynx with a dry, sterile
swab. Nasopharyngeal secretion specimens are collected by inserting a swab
with flexible shaft through the nostril to the nasopharynx.
2.Rectal swabs and stool specimens:
Used to detect rotavirus, enteric adenoviruses
and enteroviruses. Rectal swabs are collected by inserting a swab (3-5) cm into the
rectum to obtain feces.
3.Urine:
CMV, mumps, rubella, measles, polyomaviruses and adenoviruses can be
detected in urine. Virus recovery may be increased by processing multiple (2-3)
specimens because virus can be shed intermittently or in low numbers. The best
specimen is at least (10 ml) of a clean-voided first-morning urine.
4.Blood:
Used to detect CMV, HSV, enterovirus and adenoviruses. (50-10 ml) of
anticoagulated blood collected in a vacutaner tube is needed.
5.Tissue:
Useful for detecting viruses that infect lung (CMV, influenza virus,
adenovirus), brain (HSV), and gastrointestinal tract (CMV). Specimens are collected
during surgical procedures.
Specimen transport and storage:
1-All specimens collected for detection of viruses should be collected with aseptic
technique and transported immediately to the laboratory by using transport media (in
screw capped tube) and should be placed in ice, specimens for viral isolation should not
be kept at room or higher temperature. Under unusual circumstances, specimens may
need to be held for days before processing. For storage up to 5 days, hold specimen at
4
°
C. Storage for 6 or more days should be at – 20
°
C or preferably at – 70
°
C.
2-Some antibiotics (e.g penicillin, streptomycin, nystatin) should be added to viral
transport media especially when contamination with microbial flora is expected.
Examples of successful transport media include Stuart’s medium, Amie’s medium.
3-Centrifuge under refrigeration, the supernatant will contain the viruses.
4-Filter the supernatant to give crude liquid of viruses.
5-Detection of viruses in this crude liquid of viruses by 3 methods
Overview of diagnostic methods
In general, diagnostic tests can be grouped into 3 categories.: (1) direct detection,
(2) indirect examination (virus isolation), and (3) serology. In direct examination,
the clinical specimen is examined directly for the presence of virus particles, virus
antigen or viral nucleic acids. In indirect examination, the specimen into cell
culture, eggs or animals in an attempt to grow the virus: this is called virus
isolation. Serology actually constitute by far the bulk of the work of any virology
laboratory. A serological diagnosis can be made by the detection of rising titres of
antibody between acute and convalescent stages of infection, or the detection of
IgM. In general, the majority of common viral infections can be diagnosed by
serology. The specimen used for direction detection and virus isolation is very
important. A positive result from the site of disease would be of much greater
diagnostic significance than those from other sites. For example, in the case of
herpes simplex encephalitis, a positive result from the CSF or the brain would be
much greater significance than a positive result from an oral ulcer, since
reactivation of oral herpes is common during times of stress.
1. Direct Examination of Specimen
Electron Microscopy morphology / immune electron microscopy
Light microscopy histological appearance - e.g. inclusion bodies
Antigen detection immunofluorescence, ELISA etc.
Molecular techniques for the direct detection of viral genomes
2. Indirect Examination
Cell Culture - cytopathic effect, haemadsorption, confirmation by neutralization,
interference, immunofluorescence etc.
Eggs pocks on CAM - haemagglutination, inclusion bodies
Animals disease or death confirmation by neutralization
3. Serology
Detection of rising titres of antibody between acute and convalescent stages
of infection, or the detection of IgM in primary infection.
Specimen selection for viral testing depends on the suspected viral etiologies and disease syndrome. Throat and nasopharyngeal swabs are used for different viruses, while rectal swabs, urine, blood, and tissue samples are also collected for specific viral detection. Proper specimen transport and storage are crucial for accurate testing results, including using viral transport media and maintaining appropriate temperatures. Diagnostic methods include direct detection, virus isolation, and serology to detect viral particles, antigens, and nucleic acids.
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Presentation Transcript
Viral Specimen Selection and Collection: Specimen selection depends on the specific disease syndrome, viral etiologies suspected and time of year. 1.Throat, nasopharyngeal swab or aspirate: Throat swabs are acceptable for recovering enteroviruses, adenoviruses and HSV, whereas nasopharyngeal swab or aspirate specimens are preferred for the detection of influenza and parainfluenza viruses. Throat specimens are collected by rubbing inflamed, vesiculated, or purulent areas of the posterior pharynx with a dry, sterile swab. Nasopharyngeal secretion specimens are collected by inserting a swab with flexible shaft through the nostril to the nasopharynx.
2.Rectal swabs and stool specimens: Used to detect rotavirus, enteric adenoviruses and enteroviruses. Rectal swabs are collected by inserting a swab (3-5) cm into the rectum to obtain feces. 3.Urine: CMV, mumps, rubella, measles, polyomaviruses and adenoviruses can be detected in urine. Virus recovery may be increased by processing multiple (2-3) specimens because virus can be shed intermittently or in low numbers. The best specimen is at least (10 ml) of a clean-voided first-morning urine. 4.Blood: Used to detect CMV, HSV, enterovirus and adenoviruses. (50-10 ml) of anticoagulated blood collected in a vacutaner tube is needed. 5.Tissue: Useful for detecting viruses that infect lung (CMV, influenza virus, adenovirus), brain (HSV), and gastrointestinal tract (CMV). Specimens are collected during surgical procedures.
Specimen transport and storage: 1-All specimens collected for detection of viruses should be collected with aseptic technique and transported immediately to the laboratory by using transport media (in screw capped tube) and should be placed in ice, specimens for viral isolation should not be kept at room or higher temperature. Under unusual circumstances, specimens may need to be held for days before processing. For storage up to 5 days, hold specimen at 4 C. Storage for 6 or more days should be at 20 C or preferably at 70 C.
2-Some antibiotics (e.g penicillin, streptomycin, nystatin) should be added to viral transport media especially when contamination with microbial flora is expected. Examples of successful transport media include Stuart s medium, Amie s medium. 3-Centrifuge under refrigeration, the supernatant will contain the viruses. 4-Filter the supernatant to give crude liquid of viruses. 5-Detection of viruses in this crude liquid of viruses by 3 methods
Overview of diagnostic methods In general, diagnostic tests can be grouped into 3 categories.: (1) direct detection, (2) indirect examination (virus isolation), and (3) serology. In direct examination, the clinical specimen is examined directly for the presence of virus particles, virus antigen or viral nucleic acids. In indirect examination, the specimen into cell culture, eggs or animals in an attempt to grow the virus: this is called virus isolation. Serology actually constitute by far the bulk of the work of any virology laboratory. A serological diagnosis can be made by the detection of rising titres of antibody between acute and convalescent stages of infection, or the detection of IgM. In general, the majority of common viral infections can be diagnosed by serology. The specimen used for direction detection and virus isolation is very important. A positive result from the site of disease would be of much greater diagnostic significance than those from other sites. For example, in the case of herpes simplex encephalitis, a positive result from the CSF or the brain would be much greater significance than a positive result from an oral ulcer, since reactivation of oral herpes is common during times of stress.
1. Direct Examination of Specimen Electron Microscopy morphology / immune electron microscopy Light microscopy histological appearance - e.g. inclusion bodies Antigen detection immunofluorescence, ELISA etc. Molecular techniques for the direct detection of viral genomes 2. Indirect Examination Cell Culture - cytopathic effect, haemadsorption, confirmation by neutralization, interference, immunofluorescence etc. Eggs pocks on CAM - haemagglutination, inclusion bodies Animals disease or death confirmation by neutralization
3. Serology Detection of rising titres of antibody between acute and convalescent stages of infection, or the detection of IgM in primary infection. Classical Techniques 1. Complement fixation tests (CFT) 2. Haemagglutination inhibition tests 3. Immunofluorescence techniques (IF) 4. Neutralization tests 5. Single Radial Haemolysis Newer Techniques 1. Radioimmunoassay (RIA) 2. Enzyme linked immunosorbent assay (EIA) 3. Particle agglutination 4. Western Blot (WB) 5. Recombinant (RIBA), line immunoassay (Liatek) etc. immunoblot assay
