Jalava T, Kallio A, Leinonen A W, Ranki M
Orion Corporation, Orion Pharmaceutica, Helsinki, Finland.
Mol Cell Probes. 1990 Oct;4(5):341-52. doi: 10.1016/0890-8508(90)90025-u.
Nucleic acid hybridization methods in routine diagnosis of micro-organisms have been limited by the tedious assay procedures. We have previously described the sandwich hybridization method which allows convenient testing of biological specimens. In this paper we describe the adaptation of the solution hydridization method into the microtitre plate format using 35S-isotope as label. Using 3-hour hybridization followed by 2-hour collection of the hybrids a sensitivity of 5 x 10(5) target DNA molecules was achieved. The method was applied for identification of human papillomaviruses in crude gynaecological specimens. A simple 1-day assay protocol was achieved with high HPV type specificity. The specificity was confirmed by testing a variety of unrelated micro-organisms, none of which gave a positive signal in the test. Results, obtained as numerical values, were easy to interpret; positive and negative samples gave clearly distinguishable signals.
微生物常规诊断中的核酸杂交方法一直受到繁琐检测程序的限制。我们之前描述过夹心杂交方法，该方法能方便地检测生物标本。在本文中，我们描述了将溶液杂交方法改编为微量滴定板形式，使用³⁵S同位素作为标记。通过3小时杂交，随后2小时收集杂交体，实现了对5×10⁵个靶DNA分子的灵敏度。该方法用于粗制妇科标本中人乳头瘤病毒的鉴定。实现了一个简单的1天检测方案，具有高HPV型特异性。通过检测多种无关微生物确认了特异性，这些微生物在检测中均未给出阳性信号。以数值形式获得的结果易于解释；阳性和阴性样品给出明显可区分的信号。
