For detection of extremely low numbers of Chlamydia bacteria, we optimized existing real-time PCR technology at several steps from sample collection through analysis of duplex reverse-transcriptase PCR assays.
- Sampling was optimized for preservation of target nucleic acids in specimens. This is important for detection of low pathogen concentrations.
- Nucleic acid extraction was improved for maximum concentration of rare target nucleic acids such that as few as 7 copies of a nucleic acids per milliliter of original sample (blood, urine, tissue, ...) can be reliably detected.
- Chemistry and thermal cycling strategy of real-time PCR was optimized for robust amplification.
- For quantification of gene transcription, a method for simultaneous amplification of 2 target mRNAs in a single real-time PCR was developed. This method provides high accuracy, speed, sensitivity, and ease of detection and quantification of gene activity, and is also the basis of our reverse transcription PCRs for detection of RNA viruses.
- New assays can typically be developed and fully validated within 2 weeks if target nucleic acids are available. We are continuously developing new assays for diagnostic targets of interest. We are also interested in suggestions for assays of diagnostic value and encourage inquiries about such new assays. Please contact us with your questions and/or suggestions.
The combined use of modified nucleic acid extraction and real-time PCR methodology in our diagnostic platform can generate results within 3 hours. Batch nucleic acid extraction followed by real-time PCR on the same day allows communication of results within 24 hours. Finally, the robust assay technology ensures reliable results.
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