Schluger N, Godwin T, Sepkowitz K, Armstrong D, Bernard E, Rifkin M, Cerami A, Bucala R
New York Hospital-Cornell Medical Center, Manhasset 11030.
J Exp Med. 1992 Nov 1;176(5):1327-33. doi: 10.1084/jem.176.5.1327.
Pneumocystis carinii pneumonia is a leading cause of morbidity and mortality in patients with the acquired immunodeficiency syndrome (AIDS). Much remains unknown about the basic biology of P. carinii and studies of this infection have been hampered by the lack of cultivation methods. We developed a sensitive and specific assay for P. carinii by utilizing DNA amplification of the P. carinii dihydrofolate reductase (DHFR) gene. By this method, P. carinii DNA was detected in the lungs of rats with experimentally induced P. carinii pneumonia 2 wk before the onset of histopathological changes. DNA amplification analysis of serum demonstrated that by 10 wk of corticosteroid treatment, 12 of 12 (100%) infected rats had circulating DHFR DNA. P. carinii DHFR DNA also was detected in the serum of patients with AIDS and active P. carinii pneumonia (12 of 14 sera collected prospectively). Patients with advanced AIDS but without a history of P. carinii pneumonia were negative by this assay (0 of 6 sera examined). Serum polymerase chain reaction may facilitate investigations into the natural history and epidemiology of P. carinii infection, provide insight into the pathogenesis of parasite dissemination, and offer a useful, noninvasive diagnostic test for the detection of human pneumocystosis.
卡氏肺孢子虫肺炎是获得性免疫缺陷综合征(AIDS)患者发病和死亡的主要原因。关于卡氏肺孢子虫的基础生物学仍有许多未知之处,且由于缺乏培养方法,对这种感染的研究受到了阻碍。我们通过利用卡氏肺孢子虫二氢叶酸还原酶(DHFR)基因的DNA扩增,开发了一种针对卡氏肺孢子虫的灵敏且特异的检测方法。通过这种方法,在实验性诱导的卡氏肺孢子虫肺炎大鼠的肺部,在组织病理学变化出现前2周就检测到了卡氏肺孢子虫DNA。血清的DNA扩增分析表明,在皮质类固醇治疗10周时,12只受感染大鼠中有12只(100%)循环中有DHFR DNA。在患有AIDS且患有活动性卡氏肺孢子虫肺炎的患者血清中也检测到了卡氏肺孢子虫DHFR DNA(前瞻性收集的14份血清中有12份)。患有晚期AIDS但无卡氏肺孢子虫肺炎病史的患者通过该检测为阴性(检测的6份血清中有0份)。血清聚合酶链反应可能有助于对卡氏肺孢子虫感染的自然史和流行病学进行研究,深入了解寄生虫播散的发病机制,并为检测人类肺孢子虫病提供一种有用的非侵入性诊断试验。