酶联合成DNA在恶性疟诊断中的应用。

Use of enzyme-linked synthetic DNA in diagnosis of falciparum malaria.

作者信息

McLaughlin G L, Ruth J L, Jablonski E, Steketee R, Campbell G H

出版信息

Lancet. 1987 Mar 28;1(8535):714-6. doi: 10.1016/s0140-6736(87)90356-4.

Abstract

Plasmodium falciparum contains a family of 21-base-long repetitive DNA sequences in its genome. A 21-base synthetic DNA oligomer, formerly labelled with phosphorus-32 for autoradiographic detection of P falciparum DNA, was covalently coupled to alkaline phosphatase for histochemical detection. The conjugate (PFR1-AP) detected purified P falciparum DNA with a sensitivity and specificity equal to that of 32P-labelled probes after 2-day exposures. PFR1-AP did not detect host DNA or DNA of other Plasmodium species. In African blood specimens PFR1-AP specifically detected P falciparum infections of 100 parasites/microliter. This sensitive, rapid, nonisotopic probe will allow more widespread use of DNA hybridisation in the diagnosis of malaria.

摘要

恶性疟原虫基因组中含有一个由21个碱基长的重复DNA序列组成的家族。一种21个碱基的合成DNA寡聚物，以前用磷-32标记用于恶性疟原虫DNA的放射自显影检测，现在与碱性磷酸酶共价偶联用于组织化学检测。在经过2天的曝光后，这种偶联物（PFR1-AP）检测纯化的恶性疟原虫DNA的灵敏度和特异性与32P标记的探针相当。PFR1-AP未检测到宿主DNA或其他疟原虫物种的DNA。在非洲血液样本中，PFR1-AP能特异性检测出每微升含有100个疟原虫的恶性疟原虫感染。这种灵敏、快速的非同位素探针将使DNA杂交在疟疾诊断中得到更广泛的应用。