Polymerase chain reaction: trenches to benches
Polymerase chain reaction: trenches to benches
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聚合酶链式反应：从战壕到长凳
DOI：
10.1128/jcm.29.7.1281-1285.1991
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发表时间：
1991
影响因子：
9.4
通讯作者：
D. Persing
中科院分区：
文献类型：
--
作者：
D. Persing
分子诊断技术在传染病诊断中的应用是基于对病原体DNA或RNA中独特的“特征序列”的识别；在大多数情况下，在临床标本中检测到具有特征的核酸痕迹被推定为现在(或最近)感染的证据。然而，尽管幸运地引入了用于检测感染性病原体的核酸探针，但相对较少的实验室定期使用它们，主要是因为它们在技术上仍然要求很高，难以自动化，而且往往缺乏微生物标本所需的灵敏度(5、20)。因此，除了少数例外，对感染性病原体的检测仍然局限于培养确认，并没有消除对原代培养的需要。进入聚合酶链式反应(PCR)(23，24)。在过去十年里发展起来的分子生物学的基本技术中，没有一项技术产生了更大的影响。这种巧妙的方法于1985年首次被描述，它使用重复的寡核苷酸指导的DNA合成来进行目标核酸序列的体外复制，形成了一个极其敏感的系统的基础，用于扩增和检测特定的核酸序列(图1)。除了在人类遗传学和临床微生物学方面的许多已发表的应用(最近在参考文献5和20中综述)，聚合酶链式反应还提供了在实验室中完成仅仅十年前还不可能完成的任务的手段，例如从木乃伊组织中回收DNA(18)和在档案材料中鉴定人类病原体(21)。事实上，根据他们开发的扩增技术的精神，发表的用于传染病诊断的新的聚合酶链式反应应用的报告正在以似乎是指数的速度积累。然而，尽管人们对这项技术有着非凡的热情，并在其应用上投入了大量的人力和财力，但聚合酶链式反应仅在少数几个中心作为临床服务常规进行。为什么这项重要的技术在其月光构思后的8年里没有走出发展的“战壕”(16)？尽管诊断界抱怨说，对许可协议的限制阻碍了聚合酶链式反应的广泛使用，但实际上是一系列技术问题，其中一些是由技术本身造成的，阻碍了它成为临床实验室的工作台程序。这次小型审查的目的是简要总结这些问题，并说明为解决这些问题正在采取的措施。
The application of molecular diagnostic techniques to the diagnosis of infectious disease is based on the identification of unique "signature sequences" in the DNA or RNA of a pathogen; the detection of characteristic nucleic acid traces in a clinical specimen is presumed, in most cases, to be evidence of present (or recent past) infection. However, despite the auspicious introduction of nucleic acid probes for the detection of infectious agents, relatively few laboratories employ them on a regular basis, largely because they are still technically demanding and difficult to automate and often lack the sensitivity required for microbiological specimens (5, 20). Thus, with only a few exceptions, probes for infectious agents are still confined to culture confirmation and have not eliminated the need for primary culture. Enter the polymerase chain reaction (PCR) (23, 24). Of the basic techniques in molecular biology that have been developed in the last decade, none has had a greater impact. First described in 1985, this ingenious method uses repeated cycles of oligonucleotide-directed DNA synthesis to carry out in vitro replication of target nucleic acid sequences, forming the basis of an extremely sensitive system for the amplification and detection of specific nucleic acid sequences (Fig. 1). In addition to numerous published applications in human genetics and clinical microbiology (reviewed recently in references 5 and 20), PCR has provided the means to accomplish in the laboratory what only a decade ago was impossible, such as the recovery of DNA from mummified tissues (18) and the identification of human pathogens in archived material (21). Indeed, published reports of new PCR applications for the diagnosis of infectious disease, in the spirit of the amplification technology they exploit, are accumulating at a seemingly exponential rate. However, despite the extraordinary enthusiasm surrounding this technique and the considerable investment of human and financial resources in its applications, PCR is routinely performed as a clinical service in only a few centers. Why has this important technology not emerged from the developmental "trenches" in the 8 years since its moonlit conception (16)? Despite complaints from the diagnostic community that restrictions on licensing agreements have hindered the widespread use of PCR, it is in fact an array of technical problems, some created by the technique itself, that have prevented it from becoming a clinical laboratory bench procedure. The purpose of this minireview is to briefly summarize these problems and describe the measures that are being taken to address them.
HIV-1 DNA 的灵敏非同位素杂交测定。
影响因子：
2.9
作者：
Keller,GH;Huang,DP;Manak,MM
通讯作者：
Manak,MM
影响因子：
158.5
作者：
RELMAN, DA;LOUTIT, JS;TOMPKINS, LS
通讯作者：
TOMPKINS, LS
DOI：
10.1056/nejm199001183220307
发表时间：
1990-01
期刊：
The New England journal of medicine
影响因子：
--
作者：
B. Eisenstein
通讯作者：
B. Eisenstein