UNIT 8: LABORATORY DIAGNOSIS FOR ARBOVIRUSES, ROBOVIRUSES, dsRNA

LABORATORY DIAGNOSIS FOR FLAVIVIRIDAE, ALPHAVIRUS, RUBIVIRUS, and BUNYAVIRIDAE

FLAVIVIRIDAE
VIRUS VIRAL ISOLATION DIRECT DETECTION SEROLOGY

● From tissue, blood, or

CSF ● RT-PCR target: ● Japanese Encephalitis
JAPANESE
● Difficult because of ○ NS5 gene Specific IM antibodies
ENCEPHALITIS
transient and low-level ○ 5'UTR & preM gene from blood & CSF
VIRUS
viremia ○ C gene ● Antigen: E-protein
● Requires BSL-3

● RT-PCR:
○ Rapid identification and serotyping in
acute phase
● RT-PCR target:
○ Capsid and pre-membrane genes
● ELISA IM, IgG
(C/prM)
DENGUE VIRUS ● Antigen: DENV
○ NS5
Envelope proteins
○ 3'NC
○ E/NS1
○ 3'UTR / 5'UTR-C
● Non-structural protein-1 (NS-1) antigen
(surface)
● Tissue IHC
(Immunohistochemistry) & ELISA antigen
capture ● ELISA IgM antibody
YELLOW FEVER Virus recovered from blood ○ Target: Envelope and NV3 ● Antigen: E protein
VIRUS during first 4 days of illness glycoprotein
● RT-PCR
○ RT-PCR target: NS5, 5'UTR

● RT-PCR Target
● ELISA-Specific IgM & IgG
ST. LOUIS ○ NS2
antibody to SLEV
ENCEPHALITIS ○ NS3
● Antigen: Pre-membrane
VIRUS ○ NS5
and E protein

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 UNIT 8: LABORATORY DIAGNOSIS FOR ARBOVIRUSES, ROBOVIRUSES, dsRNA

● ELISA,
Immunofluorescence
● RT-PCR Target
Assay (IgG, IgM)
WEST NILE VIRUS ○ 3'UTR
● Antigen: Pre-membrane
○ 5'UTR/C
protein

● RT-PCR
○ Urine tested with serum
● ELISA IgM & IgG
○ Positive result on either test confirms
ZIKA VIRUS ○ Antigen: NS1
Zika virus infection
● RT-PCR Target:
○ NS5

ALPHAVIRUS
VIRUS VIRAL ISOLATION DIRECT DETECTION SEROLOGY

CHIKUNGUNYA ● RT-PCR Target ● ELISA

Viral Isolation or Viral
VIRUS ○ E1 ○ IgM & lg G antibodies
Culture can be done
(ALPHAVIRUS) ○ nsp1 ○ Antigen: Envelope proteins

RUBIVIRUS
VIRUS DIRECT DETECTION SEROLOGY

● RT-PCR Target:
○ N or H gene ● ELISA
○ Specimen: Swabs ○ IgM & IgG antibodies
RUBELLA VIRUS ■ Nasopharyngeal ○ Antigen: Envelope
■ Nasal glycoproteins (E1, E2)
■ Oropharyngeal

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 UNIT 8: LABORATORY DIAGNOSIS FOR ARBOVIRUSES, ROBOVIRUSES, dsRNA

BUNYAVIRIDAE
VIRUS DIRECT DETECTION SEROLOGY
ORTHOBUNYAVIRUS
(La Crosse Virus) ● Serum or CSF IgM
● RT-PCR target: M-segment polyprotein genes
● Antigen: Glycoprotein-1

● ELISA (IgM & IgG)

HANTAVIRUS ● RT-PCR target: N gene
● Antigen: nucleocapsid protein