Detection of microorganisms in vessel wall specimens of the abdominal aorta: development of a PCR assay in the absence of a gold standard.
A procedure in which the "Invitrogen Easy-DNA" kit was followed by a silica-based method for the isolation of DNA was developed for extraction of PCR-inhibitor-free DNA from up to 300 mg of human vessel wall tissue. Optimally designed PCR assays were developed for the detection of at least one infected cell in this amount of tissue. Details of the procedure are given for the detection of DNA of Chlamydia pneumoniae, cytomegalovirus and herpes simplex virus type 1 and type 2 in human vessel walls. The procedure can serve as a reference method or as a gold standard when a high-performance method is needed.
Meijer
A
A
Research Laboratory for Infectious Diseases, National Institute of Public Health, Bilthoven, The Netherlands.
van der Vliet
J A
JA
Schouls
L M
LM
de Vries
A
A
Roholl
P J
PJ
Ossewaarde
J M
JM
D001012:Aorta, Abdominal / Q000382:microbiology* / Q000821:virology; D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D004279:DNA, Viral / Q000302:isolation & purification; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004269:DNA, Bacterial; D004279:DNA, Viral; D015345:Oligonucleotide Probes