1 VIROLOGY LABS Adina Glauber

CPE
Syncytium = cell fusion → large multinucleated cells: HIV,
measles, HSV.
Lysis = cell death + membrane break: enteroviruses.
Intranuclear inclusions: papovaviruses, adenoviruses, HSV 1+2,
Intracytoplasmic inclusions: rabies virus (Negri bodies), vaccinia

Virus Genome Samples Isolation Direct detection (Ag) Serology (Ab) CPE
Enteroviruses Generally throat swab Cell culture RT-PCR Rare use for diagnosis- cell Poliovirus, Coxsackie B,
- Polioviruses RNA ss + stool Poliovirus, Coxsackie B, culture is sufficient. Echoviruses:
- Coxsackie A CSF Echoviruses - can be Neutralization test: Lytic CPE
- Coxsackie B grown → CPE. 1. Plaque assay
- Echoviruses Laboratory animals
Coxsackie A cannot
grow on cell cultures →
new-born mice
inoculation.

2. Infectivity neutralization
Sample + known antisera
3. Demonstration of virus
infectivity neutralization

Influenza RNA ss - Nasopharyngeal swab Tissue culture RT-PCR Epidemiological studies. Hemadsorption
Segmented Nasal swab Hemadsorption Method of choice to subtype From >1 blood sample:
Envelope Nasal wash Madin-Darby canine detection Collect at acute → collect 2-3
Nasal aspirates kidney cell monolayer IF weeks later.
(MDCK) Viral Ag detection (indirect) + diagnosis if:
Chicken eggs Later becomes 4 times from acute.

IF = Immunofluorescence VL=viral load

2 VIROLOGY LABS Adina Glauber

Virus Genome Samples Isolation Direct detection (Ag) Serology (Ab) CPE
HSV1 DNA ds Vesicular lesion Cell culture IF Studies Syncytium + intranuclear
HSV2 Envelope Serum Including fibroblasts → Viral Ag detection (indirect) inclusions (Cowdry bodies)
Tegument CPE PCR
Viral DNA
Most sensitive

CMV DNA ds Blood Cell culture PCR Mother→child Owl’s eye

Envelope Urine Urine/saliva in first 2 Vertical (amniotic fluid) / non- Maternal IgM + IgG → acute Intranuclear inclusions
Tegument Saliva weeks of new-born’s life vertical (serum) transmission. CMV
→ CPE IF = shell-vial assay New-born: IgM +
➔ PP65 antigenemia

EBV DNA ds Serum - PCR ELISA -

Envelope Acute infection
Tegument VCA IgG +
VCA IgM +
EBNA IgG -
Past infection
VCA IgG +
VCA IgM -
EBNA IgG +
West Nile RNA ss + Serum yes RT-PCR Main test -
Envelope CSF Confirmation test IgG + IgM
Neutralization assay

IF = Immunofluorescence VL=viral load

3 VIROLOGY LABS Adina Glauber

Virus Genome Samples Isolation Direct detection (Ag) Serology (Ab) CPE
Zika RNA ss + Serum RT-PCR IgM (Zika MAC ELISA) -
Envelope Main test Plaque reduction neutralization
test (PRNT)
Rabies RNA ss - Autopsy neurons Cell culture RT-PCR - Cytoplasmic inclusion
Envelope CPE Rapid diagnosis Negri bodies
IF
Viral Ag detection

HIV RNA ss+ Plasma Studies RT-PCR Routine -

(twice) Serum Grows slowly Qualitative Screening = ELISA = rapid
Reverse Saliva/urine (rare use) Require special lab On agar gel electrophoresis, Sensitivity ↑
transcriptase Result = +/- HIV 4th gen. → IgM, IgG, p24 Ag
Envelope Transport ASAP + 2- Quantitative
Nucleocapsi 4°c Hybridization with probes, Confirmation = Western Blot
d Spectrophotometry using Indirect IF
ELISA wells
Specificity ↑ → real + result
Result = VL
Steps:
Real-Time PCR
Molecular probes bound with
2 signal molecules:
Quencher (prevents)
Fluorescent to report.
Sanger technique
Advanced. used in subtype
determination, mutation
resistance detection,
phylogenetic analysis.
HIV p24 Ag detection

IF = Immunofluorescence VL=viral load

4 VIROLOGY LABS Adina Glauber

Virus Genome Samples Isolation Direct detection (Ag) Serology (Ab) CPE
HAV RNA ss + Serum - - Acute = IgM + -
Vaccine Naked Recovery / Vaccination = IgG +
HBV DNA Serum Lab animals Viral Ag - sandwich ELISA Window HBs Ag - -
Vaccine partially ds Chimpanzees HBe Ag -
HBV DVA -
Partially Biochemical test: ALT NOT USED
Anti HBs Ab-
circular ↑ Anti HBc IgM+
Envelope Anti HBe -
Acute HBs Ag +
HBe Ag +
HBV DNA +
Anti HBs Ab -
PCR HBc IgM+ IgG-
Qualitative Anti HBe -
Result = +/- HIV Chronic HBs Ag +
Quantitative replicative HBe Ag +
HBV DNA +
Result = VL
Anti HBs Ab -
HBc IgM- IgG+
Anti HBe -
Chronic non- HBs Ag +
replicative HBe Ag -
HBV DNA -
Anti HBs Ab -
HBc IgM- IgG+
Anti HBe -
Recovery Hbs Ag-
= cured HBe Ag-
HBV DNA -
Anti Hbs Ab+
Hbc IgG+ IgM-
HBe Ab - not relevant
Immune from HBs Ag-
natural Anti HBs Ab+
infection Anti HBc Ab+
Vaccine HBs Ag-
Anti HBs Ab+
Anti HBc Ab-
Co-infection HBs Ag+
(HBV+HDV HBs Ab-
both acute) HBe Ag+
HBc IgM+
HBc IgG-
HDV IgM+

IF = Immunofluorescence VL=viral load

5 VIROLOGY LABS Adina Glauber

Virus Genome Samples Isolation Direct detection (Ag) Serology (Ab) CPE
HCV RNA ss + Serum - RT-PCR ELISA -
Qualitative Anti-HCV (persist after recovery)
No Active viral replication. HCV RNA + = infection
Vaccine Early diagnosis in seroconversion HCV RNA - = post-infection
window period.
Quantitative Acute in HCV RNA +
Therapy monitoring. window or HCV Ab -
Chronic in
Genotype assay immunosupp.
Genotype 2/3 respond better than 1 Active HCV RNA +
in PEG-IFN + Ribavirine therapy. (acute/chronic) HCV Ab +
Sequencing Recovered HCV RNA -
Phylogenetic analysis Or VL↓ HCV Ab -
Resistant mutations detection
RFLP amplicons
Using cutting enzyme to detect
mutation sites (mutation = cutting
site misses → heavier fragment).
Uses electrophoresis.
LiPA amplicons

HPV DNA ds Cervical swab See CPE PCR - Conventional PAP / Liquid based cytology
Naked (PAP test) On glass slide for HPV DNA
or
In liquid Genotype InnoLiPA
medium Identify specific HPV type from 32
types
E6/E7 → high potential of
malignant transformation.

IF = Immunofluorescence VL=viral load

6 VIROLOGY LABS Adina Glauber

https://www.youtube.com/watch?v=KcHCBEngG9s&feature=youtu.be

IF = Immunofluorescence VL=viral load