[Development and application of quality control materials for human papillomavirus types 16 and 18 nucleic acid test]

To develop quality control materials for human papillomavirus (HPV) types 16 and 18 molecular detection and to evaluate the applicability of the materials in external quality assessment (EQA) of HPV-16, 18 clinical detection.

The target gene from HPV16 was cloned into the pEGFP-C1 plasmid, then the recombinant plasmid pEGFP-C1-HPV16 was transfected into the HPV18 carrying HeLa cells. The cultured epithelial cells carrying both HPV16 target gene and HPV18 were collected and fixed using methanol. EQA samples for HPV types 16 and 18 test diluted to the concentrations 1:10, 1:50, 1:100, 1:500 were prepared from the above prepared cells and distributed to 73 EQA participants nationwide to undergo RT-PCR. The reports from the participants were summarized and evaluated. Three samples were blindly mailed to Urumqi, Hohhot, and Xiamen respectively to observe the influence of post.

Forty-four of the 73 PCR laboratories sent feedback. For the quality control materials of the concentrations 1:10, 1:50, 1:100, and 1:500 the corresponding Ct values were 29.10, 31.19, 32.15, and 32.73 respectively for HPV-16, and 30.32, 32.13, 32.22, and 35.55 respectively for HPV-18. Stability test indicated that the quality control materials were stable at least for 40 days when stored at 4 degrees C. The EQA data from 44 participants showed, that the average fit rate was 95.1% for the high concentration positive samples and was 57.4% for the low concentration samples, and was 98.3% for the negative samples. No significant changes were detected by real-time PCR in the returned EQA samples that were blindly mailed to Urumqi, Hohhot, and Xiamen.

A quality control materials for HPV types 16 and 18 molecular detection has been developed, and quality assessment verifies its applicability in EQA.