Rapid and sensitive diagnosis of human adenovirus infections by a generic polymerase chain reaction
Rapid and sensitive diagnosis of human adenovirus infections by a generic polymerase chain reaction
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DOI：
10.1016/s0166-0934(00)00269-x
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发表时间：
2001-04-01
影响因子：
3.1
通讯作者：
Echevarría, JE
中科院分区：
文献类型：
--
作者：
Avellón, A;Pérez, P;Echevarría, JE
提出了一种新的腺病毒特异性巢式聚合酶链反应（PCR）方法。它被设计在腺病毒基因组的六联体蛋白基因内，能够在广泛的临床样本中检测所有47种人类腺病毒类型的DNA。包括一个敏感的内部控制系统，能够确保适当的分析条件，以扩增少至100个异种DNA分子，以避免假阴性结果。灵敏度估计约为每管含有插入的第一扩增产物的质粒10个分子。该方法在急性角膜结膜炎患者的31/43 / 10急性呼吸道疾病住院患者的鼻咽吸入物和2/26出血性膀胱炎患者的尿液样本中检测出腺病毒感染，敏感性优于细胞培养或免疫荧光抗原检测（IF）快速诊断呼吸道标本。F组腺病毒特异性乳胶免疫测定阳性的17例粪便中只有2例PCR阴性。内部控制系统避免了另外两个粪便样本的假阴性结果。总之，下面描述的方法被证明可用于快速诊断腺病毒感染，其灵敏度高于IF抗原检测。(C) 2001 Elsevier Science B.V.版权所有
A new adenovirus specific nested polymerase chain reaction (PCR) method is described. It was designed inside the hexon protein gene of the adenovirus genome, and was able to detect DNA of all 47 human adenovirus types in a wide range of clinical samples. A sensitive internal control system able to assure proper analytical conditions for the amplification of as few as 100 molecules of a heterologous DNA was included to avoid false negative results. Sensitivity was estimated at about 10 molecules per tube of a plasmid containing an insert of the first amplification product. The method was able to detect adenovirus infection in 31/43 conjunctival scrapings from patients with acute kerato conjunctivitis 10/40 nasopharyngeal aspirates from patients admitted to hospital with acute respiratory disease and 2/26 urine samples from patients with haemorrhagic cystitis with better sensitivity than cell culture or rapid diagnosis by antigen detection by immunofluorescence (IF) in the case of respiratory specimens. Only two of 17 stools positive for a group F adenovirus specific latex immunoassay were PCR negative. The internal control system avoided a false negative result on another two stool samples. In conclusion, the method described below was shown to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection by IF. (C) 2001 Elsevier Science B.V. All rights reserved.