Kitada K, Oka S, Kimura S, Shimada K, Serikawa T, Yamada J, Tsunoo H, Egawa K, Nakamura Y
Department of Tumor Biology, University of Tokyo, Japan.
J Clin Microbiol. 1991 Sep;29(9):1985-90. doi: 10.1128/jcm.29.9.1985-1990.1991.
Pneumocystis carinii is a eukaryotic microbe which causes fatal pneumonia in patients with AIDS. Oligonucleotide primers were used to amplify the 5S rDNA sequence of P. carinii by the polymerase chain reaction (PCR) in various clinical and animal samples. Of 35 independent lung specimens tested, PCR detected the P. carinii sequence in all 23 cases which were known to be P. carinii infected, i.e., 15 from mice, 1 from rat, 3 from human autopsy, and 4 from biopsy of AIDS patients by needle aspiration. The results were consistent with clinical and microscopic diagnosis. The detection was highly sensitive and specific. Direct sequencing of these amplified DNAs revealed homogeneity of 5S rDNA sequences of independent isolates from mice, rats, and humans. Preliminary trials manifested efficacy of the PCR method to detect P. carinii sequences in induced sputum or blood from AIDS patients, the latter case suggesting that P. carinii might enter peripheral blood via phagocytosis or direct intrusion. Development of less-invasive or noninvasive PCR diagnostic techniques to detect P. carinii infection would greatly facilitate therapeutic and prophylactic management of P. carinii pneumonia.
卡氏肺孢子菌是一种真核微生物,可导致艾滋病患者发生致命性肺炎。使用寡核苷酸引物通过聚合酶链反应(PCR)在各种临床和动物样本中扩增卡氏肺孢子菌的5S rDNA序列。在检测的35个独立肺标本中,PCR在所有23例已知感染卡氏肺孢子菌的病例中均检测到了卡氏肺孢子菌序列,即15例来自小鼠,1例来自大鼠,3例来自人体尸检,4例来自艾滋病患者经针吸活检的样本。结果与临床和显微镜诊断一致。该检测具有高度的敏感性和特异性。对这些扩增的DNA进行直接测序显示,来自小鼠、大鼠和人类的独立分离株的5S rDNA序列具有同源性。初步试验表明,PCR方法可有效检测艾滋病患者诱导痰或血液中的卡氏肺孢子菌序列,后一种情况表明卡氏肺孢子菌可能通过吞噬作用或直接侵入进入外周血。开发用于检测卡氏肺孢子菌感染的侵入性较小或非侵入性的PCR诊断技术将极大地促进卡氏肺孢子菌肺炎的治疗和预防管理。