Wakefield A E, Pixley F J, Banerji S, Sinclair K, Miller R F, Moxon E R, Hopkin J M
Molecular Infectious Diseases Group, John Radcliffe Hospital, Oxford, UK.
Lancet. 1990 Aug 25;336(8713):451-3. doi: 10.1016/0140-6736(90)92008-6.
Oligonucleotide primers and probes were used in the polymerase chain reaction to amplify Pneumocystis carinii specific DNA sequences from alveolar lavage samples from 47 diagnostic bronchoscopies. No P carinii DNA was found in lavage from 10 immunocompetent patients; only low levels were found in 3 of 13 samples from immunosuppressed individuals without P carinii pneumonia (PCP), and the highest levels, readily demonstrated by simple ethidium bromide staining, were found in all of 16 samples from immunosuppressed patients with PCP confirmed by means of standard silver staining and in 4 from patients with clinical PCP but negative silver staining. DNA amplification provides a highly sensitive and specific technique for the identification of P carinii that should be valuable in epidemiological studies on this parasitic infection and in diagnosis.
使用寡核苷酸引物和探针通过聚合酶链反应从47例诊断性支气管镜检查的肺泡灌洗样本中扩增卡氏肺孢子虫特异性DNA序列。10例免疫功能正常患者的灌洗样本中未发现卡氏肺孢子虫DNA;13例无卡氏肺孢子虫肺炎(PCP)的免疫抑制个体的样本中,只有3例发现低水平的卡氏肺孢子虫DNA,而通过简单的溴化乙锭染色即可轻松显示的最高水平,在所有16例经标准银染色确诊为PCP的免疫抑制患者的样本中以及4例临床诊断为PCP但银染色阴性的患者样本中均有发现。DNA扩增为卡氏肺孢子虫的鉴定提供了一种高度灵敏且特异的技术,这在关于这种寄生虫感染的流行病学研究和诊断中应具有重要价值。