Toward the goal of reducing diagnostic false positives while retaining high sensitivity, a closed-tube nested PCR procedure has been developed for detecting low-copy-number human immunodeficiency virus (HIV) gag target DNA sequences. Master mix for amplification 2, in a hanging gel matrix at the reaction tube top, remains sequestered from the reaction space of the tube during amplification 1. A...

background: toll like receptors (tlrs) are one of the main components of the innate immune system. it has been reported that expression of these receptors are altered in the female reproductive tract (frt) during menstrual cycle. here we used a fallopian tube epithelial cell line (oe-e6/e7) to evaluate the effect of two sex hormones in modulating tlr expression. materials and methods: in this e...

We present a protocol for effi cient amplifi cation of a large number of DNA targets using a single-tube polymerase chain reaction (PCR) based on PCR suppression. This method allows amplifi cation of each DNA target with only one target-specifi c primer whereas the other primer is common for all targets. Thus, this approach requires n+1 primers for n targets instead of 2n in conventional PCR an...

BACKGROUND Homogeneous PCR methods for genotyping usually require fluorescently labeled oligonucleotide probes. Amplicon melting with the DNA dye LCGreen I was recently introduced as a closed-tube method of genotyping that does not require probes or real-time PCR. However, some single-nucleotide polymorphisms (SNPs) could not be completely genotyped without addition of a known genotype, and hig...

The PCR diagnosis of toxoplasmosis suffers from lack of standardization. Interlaboratory comparative studies of PCR methods have been performed, but intralaboratory comparisons are scarce. Here, we optimized and compared the technical performances of two PCR primer systems widely used for Toxoplasma detection. The differences between the two methods were visible only at low parasite concentrati...

Using dissociation and enhancement time-resolved lanthanide fluorometry, we have developed a quantitative competitive (QC)-PCR for measuring parathyroid hormone-related protein (PTHrP) mRNA after reverse transcription. A cloned PTHrP cDNA target was also modified by deletion of 10 bp and insertion of 21 bp in the midregion of the fragment and cloned for use as a competitor (i.e., internal stand...

A multiple-tubes procedure is described for using PCR to determine the genotype of a very small DNA sample. The procedure involves dividing the sample among several tubes, then amplifying and typing the contents of each tube separately. The results are analyzed by a statistical procedure which determines whether a genotype can be conclusively assigned to the DNA sample. Simulation studies show ...

Background: In an ectopic pregnancy (EP) the egg does not reach to the uterus, but instead implants somewhere outside it, usually in fallopian tube. In most cases the exact cause of an ectopic pregnancy is unknown but it is often a result of some sort of damage to the fallopian tube. The tube may have become blocked or narrowed by previous surgery or infection. An active immune system need to r...

Here we present a one-tube nested PCR test, which allows the detection of minimal quantities of Chlamydia trachomatis in human fluids. This assay includes the use of an internal control to avoid false negative results due to the presence of inhibitors. The results obtained show that this assay is robust enough to be used for clinical diagnosis.