Previously we described a PCR protocol for detecting the inversion in the factor VIII gene, which is a common cause of Hemophilia A. This PCR assay is challenging due to the size of the amplification (10-12 kb), the varying GC content (30%-80%) and the multiplex PCR products involved (four for carrier female). Efficient amplification of the four segments depends on three unusual modifications t...

In a recent review, Drs. Wong and Medrano provided a good overview of the problems regarding mRNA quantification using real-time PCR (1). They claim that there are several pitfalls using this method for quantification and indeed it is, without a doubt, that fluctuation during PCR will affect quantification most since minute differences in PCR efficiency greatly affect the yield. However, as the...

For detection of respiratory syncytial virus (RSV), the BD Directigen RSV rapid antigen assay was compared to Cepheid's real-time reverse transcriptase PCR RSV analyte-specific reagents. The Directigen RSV assay resulted in a 23% false-negative rate, using PCR and chart review as the gold standard, indicating that rapid RSV PCR results would be advantageous.

background and objective: rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among non-vaccinated infants for whom pertussis is life- threatening. materials and methods: targets is481 , is1001, bp0026 and human gapdh gene were used to develop a multiplex real- time pcr assay based on the ...

The use of direct nested PCR enables the detection of Plasmodium spp. from blood samples collected on filter papers without requiring the time-consuming procedures associated with DNA extraction. Direct PCR provides a rapid, highly sensitive, and cost-effective alternative to diagnosing malaria using filter paper samples and standard nested PCR.

Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular epidemiological study. Repetitive extragenic palindromic PCR (rep-PCR) is less time-consuming and more suitable for analyzing large numbers of bacterial strains in human populations. PFGE and rep-PCR provide comparable genotyping results for investigating Streptococcus mutans diversity and transmission.

accurate and rapid diagnosis of human cytomegalovirus (hcmv) disease in immunocompromised patients has remained as a challenge. quantitative competitive pcr (qc-pcr) methods for detection of hcmv in these individuals have improved the positive and negative predictive values of pcr for diagnosis of hcmv disease. in this study we used qc-pcr assay, using a co-amplified dna standard, to quantitate...

In the last decade, genome-wide gene expression data has been collected from a large number of cancer specimens. In many studies utilizing either microarray-based or knowledge-based gene expression profiling, both the validation of candidate genes and the identification and inclusion of biomarkers in prognosis-modeling has employed real-time quantitative PCR on reverse transcribed mRNA (qRT-PCR...

Peptide nucleic acid (PNA)-mediated PCR clamping (H. Orum, P. E. Nielsen, M. Egholm, R. H. Berg, O. Buchardt, and C. Stanley, Nucleic Acids Res. 21:5332-5336, 1993) was introduced as a novel procedure to selectively amplify ribosomal DNAs (rDNAs) which are not frequently found in clone libraries generated by standard PCR from complex microbial consortia. Three different PNA molecules were used;...