Inhibitory effects of urine on the polymerase chain reaction for cytomegalovirus DNA.
TLDR
The sensitivity and specificity of PCR for detecting cytomegalovirus DNA in urine were further improved by using "nested" primers and a modified PCR protocol entailing the use of reduced reactants in the first 20 cycles of a two-stage 50 cycle PCR.Abstract:
The inhibitory effects of urine samples taken from neonates and older children, some of which were known to be infected with cytomegalovirus, on the polymerase chain reaction (PCR) were investigated. Urea was the major inhibitory component of urine and inhibited the PCR at a concentration of more than 50 mM. Urine samples from older children were more inhibitory than those from neonates. This correlated with the higher concentration of urea generally found in urine samples from older children compared with neonatal urines. Two of 13 neonatal urine samples, however, were inhibitory despite low urea concentrations--presumably due to metabolites derived from parenteral nutrition. The inhibitory effects of urine were effectively removed by simple dialysis or ultrafiltration. The sensitivity and specificity of PCR for detecting cytomegalovirus DNA in urine were further improved by using "nested" primers and a modified PCR protocol entailing the use of reduced reactants in the first 20 cycles of a two-stage 50 cycle PCR.read more
Citations
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PCR inhibitors - occurrence, properties and removal.
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Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Common Strains of Escherichia coli
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Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
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Sergio Stagno,Robert F. Pass,Meyer E. Dworsky,Ronald E. Henderson,Ernest G. Moore,Philip D. Walton,Charles A. Alford +6 more
TL;DR: It is concluded that congenital cytomegalovirus infection resulting from primary maternal infection is more likely to be serious than that resulting from recurrent infection, and is morelikely to occur in upper socioeconomic groups.
Journal ArticleDOI
Detection of Cytomegalovirus in Urine from Newborns by Using Polymerase Chain Reaction DNA Amplification
TL;DR: PCR amplification may be a valuable tool for diagnosing congenital CMV infection in tissue culture and in urine specimens from newborns.