Immunochemiluminescent Southern blot assay for polymerase chain reaction detection of human parvovirus B19 DNA.
Human parvovirus B19 is not only an acute self-limited infection causing erythema infectiosum, transient aplastic crisis, foetal hydrops and arthritis but can also be a chronic infection causing chronic anaemia and associated with chronic neuropathy and vasculitis. Serologic studies have proven to be the most sensitive way to detect acute infection in the immunologically normal patient while polymerase chain reaction (PCR) assays for B19 DNA are the most sensitive way to detect chronic infection. The ability to detect B19 in clinical specimens can be further increased with a second amplification step using nested primers. However, nested PCR is both time consuming and enhances the risk of false-positive results due to contaminating DNA. In this study, we developed a sensitive immunochemiluminescent Southern blot assay for detecting PCR amplified B19 DNA with a digoxigenin labelled primer. The sensitivity and specificity of this assay were comparable to nested PCR and at least 100-fold more sensitive than a single PCR amplification.
Respiratory and Enterovirus Branch, National Center for Infectious Diseases, Atlanta, GA 30333.
E L
EL
Durigon
D D
DD
Erdman
B C
BC
Anderson
B P
BP
Holloway
L J
LJ
Anderson
D015139:Blotting, Southern / Q000379:methods*; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics*; D016731:Erythema Infectiosum / Q000175:diagnosis / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008163:Luminescent Measurements; D016732:Parvovirus B19, Human / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods
D004279:DNA, Viral