Acrongenomics Inc. recently conducted a clinical validation study at EuroGENET Laboratories SA to further establish its Nano-JETA(TM) technology platform in regards to Real Time PCR application, the Nano-JETA(TM) Real Time PCR.

The objectives of the study were:

a) to validate the Nano-JETA(TM) Real Time PCR via the detection of Ep-CAM gene and beta-actin (internal control) in peripheral blood of patients suffering from breast and colon cancer (patients were histologically and cytologically verified), and

b) to evaluate the Nano-JETA(TM) technology platform in comparison to conventional Real Time PCR methodology for Ep-CAM and beta-actin respectively in regards to stability, sensitivity, specificity and reproducibility.

The study was designed intentionally incorporating the most difficult parameters such as blind control, multi-centre clinical validation study, with variant and unidentified patient status (primary or metastatic status, follow up or on treatment).

The original number of patient samples was 250 but according to EuroGENET Labs Quality Control acceptance criteria, only 236 patient samples qualified as appropriate for participating in the study. The remaining 14 samples were rejected. Therefore, RNA was extracted from fresh blood samples of 236 patients suffering from breast and colon cancer. cDNA was produced according to both Nano-JETA(TM) and conventional cDNA protocols.

Conventional Real Time PCR and Nano-JETA(TM) Real Time PCR were performed for Ep-CAM and beta-actin. The average cycles required for Real Time PCR and Nano-JETA(TM) Real Time PCR completion were 45 and 13 respectively. Furthermore, the total reaction volume required for Real Time PCR and Nano-JETA(TM) Real Time PCR was 25microlitres and 8microlitres respectively.

Research results revealed that in the case of beta-actin (internal control) both protocols exhibited 100% specificity. Beta-actin was detectable from the 16th to 18th cycle when using common Real-Time PCR, while only 1 single cycle was required with Nano-JETA(TM) Real-Time PCR.

In regards to sensitivity, common Real-Time PCR detected 10 to the 4th cancer cells, while requiring 22 to 32 cycles. In contrast, Acrongenomics' Nano-JETA(TM) Real-Time PCR exhibited sensitivity of 10 to the 2nd cancer cells, with the use of 1 single cycle.

By using common Real-Time PCR, Ep-CAM was detected in 28 samples out of 236 (11.9% positive) and it was not found in 208 samples (88.1% negative) showing sensitivity of 10 to the 4th cancer cells as noted before.

On the other hand, by using Nano-JETA(TM) Real-Time PCR, Ep-CAM was detected in those same 28 samples -- but it was also detected in 22 additional samples (21.2%). It was not detected in 186 samples out of 236 (78.8% negative), exhibiting sensitivity of 10 to the 2nd cancer cells. This was due to the fact that Nano-JETA(TM) Real-Time PCR is one hundred times more sensitive than conventional Real Time PCR.

Acrongenomics believes that the capabilities and performance of the Nano-JETA(TM) technology platform developed by Acrongenomics Inc. could be regarded as a breakthrough application in the diagnostic market for stability, sensitivity, specificity and reproducibility.