Wolf H, Leser U, Haus M, Gu S Y, Pathmanathan R
J Virol Methods. 1986 Apr;13(1):1-8. doi: 10.1016/0166-0934(86)90066-2.
Nucleic acid hybridization is widely used for scientific applications but essentially restricted to specialized laboratories. The use of recombinant m 13 phages as hybridization probes (Hu and Messing (1980) Gene 17, 271; Messing (1983) Methods Enzymol. 101, 20) offers a considerable advantage over the commonly used recombinant plasmids as the preparation of the DNA probe is very simple and it can easily be labeled directly, e.g. with isotopes with long half-life like 125I (Commerford (1971) Biochemistry 10, 11 (1983); Gu et al. (1983) Cancer (China) 2, 129; Han and Harding (1983) Nucleic Acids Res. 11, 14) and used for hybridization. However, as the application of nucleic acid hybridization for diagnostic and epidemiological purposes becomes almost unavoidable, the logistic problems of keeping numerous individually labeled hybridization probes increase considerably and may reach prohibitory levels in less well-equipped laboratories. In a new sandwich technique, the first step involves hybridization with an unlabeled recombinant m 13 DNA carrying an insert of the desired specificity. In a second step a universally usable labeled probe directed against the m 13 part of the recombinant phage DNA is applied. This reduces considerably the problems of preparing and keeping multiple labeled probes in stock.
核酸杂交在科学应用中广泛使用，但基本上局限于专业实验室。使用重组M13噬菌体作为杂交探针（Hu和Messing（1980年）《基因》第17卷，271页；Messing（1983年）《酶学方法》第101卷，20页）与常用的重组质粒相比具有相当大的优势，因为DNA探针的制备非常简单，并且可以很容易地直接进行标记，例如用半衰期长的同位素如125I（Commerford（1971年）《生物化学》第10卷，11页（1983年）；顾等人（1983年）《癌症（中国）》第2卷，129页；Han和Harding（1983年）《核酸研究》第11卷，14页）并用于杂交。然而，随着核酸杂交在诊断和流行病学目的中的应用几乎变得不可避免，保存大量单独标记的杂交探针的后勤问题大幅增加，在设备较差的实验室中可能达到难以承受的程度。在一种新的夹心技术中，第一步涉及与携带具有所需特异性插入片段的未标记重组M13 DNA杂交。在第二步中，应用针对重组噬菌体DNA的M13部分的通用标记探针。这大大减少了制备和保存多种标记探针库存的问题。
