Detection of sickle cell beta S-globin allele by hybridization with synthetic oligonucleotides.

Abstract

Two 19-base-long oligonucleotides were synthesized, one complementary to the normal human j-globin gene (pA) and one complementary to the sickle cell (-globin gene (PS).The nonadecanucleotides were radioactively labeled and used as probes in DNA hybridization. Under appropriate hybridization conditions, these probes can be used to distinguish the pJA gene from the 3ss allele. The DNA from individuals homozygous for the normal fi-globin gene (fApA) only hybridized with the plA specific probe; the DNA from those homozygous for the sickle cell ,3-globin gene ((s.(s) only hybridized with the SsS specific probe. The DNA from heterozygous individuals (pATS) hybridized with both probes. This allele-specific hybridization behavior of oligonucleotides provides a general method for diagnosis of any genetic disease which involves a point mutation in the DNA sequence of a single-copy gene.

Genetic and Epidemiological Studies of Dengue Type 2 Viruses by Hybridization Using Synthetic Deoxyoligonucleotides as Probes

“…This technique has been used previously in analyses of 4~X-174 bacteriophage DNA (Wallace et al, 1979), rabbit beta-globulin DNA , sickle cell betas-globulin allele (Conner et al, 1983), mutations in codon 61 of the human N-ras gene (Bos et al, 1984), and in the diagnosis of enterotoxogenic Escherichia coli (Hill et al, 1985). As in oligonucleotide fingerprinting, hybridization with short probes can detect a change in one or more bases (Wallace et al, 1979).…”

Detection of single nucleotide polymorphisms by minisequencing on a polypyrrole DNA chip designed for medical diagnosis

“…17 Probes to be grafted (Table 1) are synthesized ex situ and purified. They are labeled at their 5 0 -end with a pyrrole residue using pyrrole-phosphoramidite building blocks and tailed with a poly(T) 10 spacer arm. The pyrrole group enables probe fixation on the gold electrodes using an electrocopolymerization process as described previously.…”

“…9 High-density oligonucleotide arrays such as variation detection arrays manufactured by Affymetrix have been developed to analyze single nucleotide variations using the principle of allelespecific oligonucleotide (ASO) hybridization. 10 Hybridization chips contain thousands of oligonucleotides, matching normal or mutant sequences. Under controlled hybridization conditions, probe-target heteroduplexes could be discriminated from perfectly matched probe-target duplexes, thus allowing the analysis of several polymorphisms or mutations.…”

Oligonucleotide Probes Complementary to Variable Regions of Ribosomal RNA Discriminate between Mycoplasma Species

“…(2) The high number of rRNA target molecules results in a sensitivity at least 100 times greater than that of bacterial DNA targets (Falkow, 1985;Tompkins et al, 1985;Eisenstein & Engleberg, 1986). (3) The use of short probes substantially reduces hybridization times and allows exact prediction of the hybridization conditions (Conner et al, 1983).…”

“…2), following the suppliers' recommendations (Biihringer Mannheim ; BRL; Pharmacia). The resulting fragments were separated on a 1 % (w/v) agarose gel, blotted onto nylon filters (Pall) and hybridized to the [~~~PIATP-end-labelled oligonucleotide probes (Wallace et al, 1979(Wallace et al, , 1981Conner et al, 1983).…”