Highly sensitive chemiluminescent method for the detection of cell contamination

✍ Scribed by Sirchia, S. M. ;Garagiola, I. ;Andreis, C. De ;Pedranzini, L. ;Poli, F. ;Grati, F. R. ;Diomelli, B. ;Simoni, G.

• Publisher: John Wiley and Sons
• Year: 1998
• Weight: 103 KB
• Volume: 13
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/(sici)1099-1271(1998090)13:5<303::aid-bio492>3.0.co;2-a

✦ Synopsis

Minisatellite analysis is commonly used in forensic disputes but can also be applied to the investigation of cell contamination. Such a problem arises, for example, when transplantation is performed. The presence of contamination has been investigated by other authors using radioactive methods. In the present study we describe a method that allows the detection of contamination with high sensitivity without using radioactive substances. Our technique is based on the use of polymerase chain reaction (PCR) ampli®cation of minisatellite sequences (VNTR), followed by chemiluminescent detection. In particular, biotin-labelled dCTP is included in the PCR mixture and detection of PCR products is obtained following the CSPD chemiluminescent protocol (Southern-Light Nucleic Acid Detection Systems). We applied this method to arti®cial mixes of DNA of two individuals with alleles of different sizes. We performed progressive dilutions of an individual DNA into the other's DNA and revealed a contamination of 1 in 2500 cells. We also tested our technique searching for maternal contamination in cord blood samples in 60 cases and revealed a 18.3% contamination. The technique that we set up proves to be a very sensitive one which could be applied not only to the detection of maternal cells in cord blood but also in studying any other kind of contamination.