A new method for direct analysis of polymerase chain reaction-amplified human papillomavirus using DNA enzyme immunoassay

1994 ◽  
Vol 24 (4) ◽  
pp. 223-226 ◽  
Author(s):  
F. Sangiuolo ◽  
L. De Santis ◽  
A. Cavicchini ◽  
U. Angeloni ◽  
C. Romanini ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Papillomavirus ◽  
Enzyme Immunoassay ◽  
Direct Analysis ◽  
New Method ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Dna Enzyme Immunoassay

High-throughput polymerase chain reaction chemiluminescent enzyme immunoassay for typing and quantifying human papillomavirus DNAs

2004 ◽  
Vol 332 (2) ◽  
pp. 349-357 ◽  
Author(s):  
Simone Ambretti ◽  
Mara Mirasoli ◽  
Simona Venturoli ◽  
Marialuisa Zerbini ◽  
Mario Baraldini ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Papillomavirus ◽  
Enzyme Immunoassay ◽  
High Throughput ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chemiluminescent Enzyme Immunoassay

Simplified reverse transcription polymerase chain reaction procedure with detection by microplate hybridization for routine screening of hepatitis A virus

1998 ◽  
Vol 44 (3) ◽  
pp. 298-302 ◽  
Author(s):  
Charlotte Arnal ◽  
Virginie Ferré-Aubineau ◽  
Bernard Besse ◽  
Sylviane Billaudel
Keyword(s):  
Polymerase Chain Reaction ◽  
Enzyme Immunoassay ◽  
Reverse Transcription ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Single Step ◽  
Chain Reaction ◽  
Complete Automation ◽  
Polymerase Chain ◽  
Dna Enzyme Immunoassay

Reverse transcription polymerase chain reaction, using either nested or seminested primers, is used extensively for the detection of viruses in small quantities. However, existing methods are prone to false positive reactions. We report here an improved polymerase chain reaction technique based on the use of longer primers (39 nucleotides) with single-step amplification, applied to the detection of hepatitis A in low quantities. While the sensitivity of this technique (10 x the 50% tissue culture infective dose) is equivalent to that of existing methods, it is a simpler procedure, less time consuming, and less susceptible to contamination and therefore provides a more reliable tool for routine diagnosis. Finally, the development of a DNA enzyme immunoassay detection technique and the complete automation of the procedure allow a large number of samples to be processed in clinical laboratories.Key words: long primers, hepatitis A virus, reverse transcription polymerase chain reaction, seminested PCR, DNA enzyme immunoassay.

DNA enzyme immunoassay: general method for detecting products of polymerase chain reaction

1991 ◽  
Vol 37 (3) ◽  
pp. 422-429 ◽  
Author(s):  
Giovanni Mantero ◽  
Antoneil Zonaro ◽  
Aiberto Albertini ◽  
Alberto Paola ◽  
Daniele Primi
Keyword(s):  
Monoclonal Antibody ◽  
Polymerase Chain Reaction ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Colorimetric Detection ◽  
Colorimetric Method ◽  
Hbv Dna ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Dna Enzyme Immunoassay

Abstract We developed a new colorimetric method, DNA enzyme immunoassay (DEIA), for detecting specific hybrids of complementary nucleic acids and applied it to the detection of hepatitis B virus (HBV) DNA amplified from serum samples by means of the polymerase chain reaction (PCR) technique. The method is based on the ability of an anti-DNA monoclonal antibody to discriminate between single-stranded and double-stranded DNA. A solid phase was coated with a specific oligonucleotide probe, internal to the amplified region of HBV DNA, via an avidin-biotin bridge. The denatured PCR product was hybridized with the solid-phase probe, and the amplified DNA probe hybrid was then incubated with a monoclonal antibody specific for double- but not single-stranded DNA. Colorimetric detection of the DNA-antibody complex was achieved by adding an anti-mouse Ig antibody labeled with horseradish peroxidase. The combined use of DEIA and PCR can reveal a few HBV genome copies present in a serum sample. This method has several advantages: (a) the sensitivity is adequate for the detection of amplified DNA; (b) the signal is associated with the hybridization event, independently of modifications of the probe or of the amplification primers; and (c) the test is simple and rapid and, most importantly, requires only the standard facilities of a routine clinical laboratory.

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