DNA amplification for the in vitro detection of Candida albicans in head and neck squamous cell carcinomas.
DNA was extracted from whole cells of Candida albicans and digested with HindIII restriction enzyme. After electrophoresis in a segment of the lane containing between 800 and 1200 base pairs (bp) of DNA fragments, a 1.1-kilobase (kb) fragment was found that hybridizes to biopsied tumor cells from head and neck squamous cell carcinomas (SCC). From the nucleotide sequence of the putative gene locus, primers were synthesized for use in a polymerase chain reaction (PCR) with DNA extracted from 18 SCC of the upper aerodigestive tract. After 30 cycles of amplification all tumors were found to contain sufficient amplified DNA to be detected in polyacrylamide or agarose gels. In contrast, template DNA from lymph nodes and malignant lymphomas failed to generate positive signals under these conditions. However, samples of DNA obtained from head and neck SCC cells in vitro, Candida glabrata, and Candida parapsilosis after PCR were found to contain homologous sequences. Application of this technique to head and neck SCC biopsies may help to identify quickly the presence of concurrent candidal species.
Werner
J A
JA
Department of Otorhinolaryngology, Head and Neck Surgery, University of Kiel, Germany.
Görögh
T
T
Lippert
B M
BM
Rudert
H
H
D001482:Base Composition; D001483:Base Sequence; D002175:Candida / Q000145:classification / Q000235:genetics; D002176:Candida albicans / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000382:microbiology*; D002874:Chromosome Mapping; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005784:Gene Amplification; D006258:Head and Neck Neoplasms / Q000382:microbiology*; D006801:Humans; D008198:Lymph Nodes / Q000378:metabolism; D008223:Lymphoma / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013698:Templates, Genetic
D004271:DNA, Fungal