The Study on Detecting Fluorescence of Hybridization Between Human Cytomegalovirus Gene in Infants and Molecular Beacons.

Abstract

Objective To design molecular beacon of human cytomegalovirus (HCMV) gene and its amplification system and find the detecting way by molecular beacon chip. Methods From June 1, 2008 to July 30, 2008, clinical positive human cytomegalovirus blood samples were collected from the following 4 hospitals, Southwest Hospital, Third Military Medical University (n=7), Chongqing Family Planning Hospital (n=8), Xinqiao Hospital of the Third Military Medical University(n=5), and Children's Hospital of Chongqing (n=9). A total of 29 cases were involved into infected group. At the same time, the clinical negative samples were collected from the same 4 hospitals, including Southwest Hospital, Third Military Medical University (n=5), Chongqing Family Planning Hospital (n=4), Xinqiao Hospital of the Third Military Medical University (n=4) and Children's Hospital of Chongqing(n=3). There were 16 cases were classified into the control. The points without probe (n=10) were designed in blank control group. The research procedures were fitted for the ethical standard formulated by the investigation in human beings committee of Southwest Hospital, Third Military Medical University. Informed consent was obtained from all participations. The software of Beacon designer was used to design the molecular beacon and molecular beacon with single extending arm of human cytomegalovirus gene. The amplification system and molecular beacon chip, and Fluorescence microscope and image analysis software were used to detect the fluorescent light and make a qualitative judgment. Results The difference between PCR products from infected group and control group was obvious in detecting the fluorescent light by the Fluorescence microscope. The fluorescent light was obviously different between 29 cases from infected group and 16 cases from control group (P<0.05). Conclusion The technology of molecular beacon chip is a technology with characteristics of high sensitivity, quick and batch in detecting infant human cytomegalovirus gene. It has many advantages such as stronger fluorescent light recognition and amplification and more accurate in signal detection. Key words: molecular beacon; human cytomegalovirus (HCMV); fluorescence microscope; infant