Atzori C, Angeli E, Agostoni F, Mainini A, Micheli V, Cargnel A
II Department of Infectious Diseases, L. Sacco Hospital, Milan, Italy.
Int J Infect Dis. 1998;3(2):76-81. doi: 10.1016/s1201-9712(99)90013-9.
To verify the clinical value of two different polymerase chain reactions (PCRs) for noninvasive diagnosis and follow-up during Pneumocystis carinii f. sp. hominis pneumonia (PCP) and to analyze the P. carinii f. sp. hominis genotypes involved.
Internal transcribed spacers (ITSs) nested PCR was applied to 630 samples (bronchoalveolar lavage, sera, peripheral blood mononuclear cells, and oropharyngeal samples) from 122 patients with acquired immunodeficiency syndrome and pneumonia and 40 control samples from 20 subjects seronegative for human immunodeficiency virus. One hundred and eighty samples also were examined by mt-rRNA PCR. Bronchoalveolar lavage samples and 33 sera were analyzed by type-specific oligonucleotide hybridization.
On bronchoalveolar lavage samples, the two PCRs consistently confirmed the morphologic diagnosis of PCP. The sensitivity of ITSs nested PCR versus mt-rRNA PCR was 57.3% versus 14.3% on sera, 32.3% versus 22. 8% on peripheral blood mononuclear cells, and 69.1% versus 48.6% on oropharyngeal samples (garglings). Both PCRs had 100% specificity. Type-specific oligonucleotide hybridization revealed in 72.2% of bronchoalveolar lavage samples a single P. carinii f. sp. hominis genotype, whereas in 27.8% co-infection with more than one strain was detected.
On noninvasive samples, ITSs nested PCR was more sensitive than mt-rRNA PCR, and it confirmed the diagnosis in all patients with PCP. For each patient with PCP at least one noninvasive sample was positive for P. carinii f. sp. hominis DNA.
验证两种不同的聚合酶链反应（PCR）在卡氏肺孢子虫肺炎（PCP）无创诊断及随访中的临床价值，并分析所涉及的卡氏肺孢子虫基因型。
对122例获得性免疫缺陷综合征合并肺炎患者的630份样本（支气管肺泡灌洗样本、血清、外周血单个核细胞和口咽样本）以及20例人类免疫缺陷病毒血清学阴性受试者的40份对照样本进行内转录间隔区（ITS）巢式PCR检测。180份样本同时进行线粒体核糖体RNA（mt-rRNA）PCR检测。采用型特异性寡核苷酸杂交法对支气管肺泡灌洗样本和33份血清进行分析。
在支气管肺泡灌洗样本中，两种PCR均一致证实了PCP的形态学诊断。血清样本中，ITS巢式PCR与mt-rRNA PCR的敏感性分别为57.3%和14.3%；外周血单个核细胞样本中分别为32.3%和22.8%；口咽样本（漱口液）中分别为69.1%和48.6%。两种PCR特异性均为100%。型特异性寡核苷酸杂交显示，72.2%的支气管肺泡灌洗样本中仅有一种卡氏肺孢子虫基因型，而27.8%的样本检测到多种菌株共感染。
在无创样本中，ITS巢式PCR比mt-rRNA PCR更敏感，且能确诊所有PCP患者。对于每例PCP患者，至少有一份无创样本卡氏肺孢子虫DNA呈阳性。
