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在血液系统肿瘤相关肺部并发症中利用支气管肺泡灌洗细胞对人巨细胞病毒进行定量分析。

Quantification of human cytomegalovirus using bronchoalveolar lavage cells in pulmonary complications associated with hematologic neoplasia.

作者信息

Ohyashiki Junko H, Nagate Akira, Ojima Tomoko, Abe Kenji, Yamamoto Kohtaro, Ohyashiki Kazuma

机构信息

Intractable Diseases Research Center, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023, Japan.

出版信息

Int J Mol Med. 2003 Jun;11(6):779-83.

Abstract

Human cytomegalovirus (CMV) has been recognized as a frequent pathogen involved in interstitial pneumonia (IP), and CMV-IP is a severe and life-threatening complication in the immunocompromised patients. The use of real-time PCR in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting a wide variety of templates including viruses. Therefore, we developed a rapid quantification system of CMV using a LightCycler in order to clarify the possible role of CMV reactivation in patients with hematologic neoplasia showing pulmonary complications. Sixty-nine bronchoalveolar lavage fluid (BALF) specimens were obtained from consecutively treated patients showing interstitial shadow including 20 patients with hematologic neoplasia. First, we determined the viral burden in BAL cells from healthy volunteers, idiopathic interstitial pneumonia (IIP) and sarcoidosis. CMV copy numbers in samples obtained from healthy volunteers, IIP and sarcoidosis, were less than 10(2) copies per 1 microg of DNA, whether or not BAL cells were composed of high percentage of lymphocytes. Among 20 patients with hematologic neoplasia analyzed, two specimens obtained from leukemia patients with pulmonary alveolar proteinosis, two from GvHD, one with CMV interstitial pneumonia and one with Hodgkin's disease had high level of CMV viral DNA. Our results suggest that measurement of CMV genomes in BAL cells using real-time PCR may be useful not only to understand the involvement of CMV in systematic respiratory tract disease but also in management of the care of respiratory complications in hematologic neoplasia.

摘要

人巨细胞病毒(CMV)已被公认为是间质性肺炎(IP)中常见的病原体,CMV-IP是免疫功能低下患者严重的、危及生命的并发症。实时PCR在分子诊断中的应用已增加到现在被公认为检测包括病毒在内的多种模板的金标准。因此,我们开发了一种使用LightCycler的CMV快速定量系统,以阐明CMV再激活在出现肺部并发症的血液系统肿瘤患者中的可能作用。从连续接受治疗且有间质性阴影的患者中获取了69份支气管肺泡灌洗液(BALF)标本,其中包括20例血液系统肿瘤患者。首先,我们测定了健康志愿者、特发性间质性肺炎(IIP)和结节病患者BAL细胞中的病毒载量。无论BAL细胞是否由高比例淋巴细胞组成,从健康志愿者、IIP和结节病患者获得的样本中,CMV拷贝数均低于每1微克DNA 10²拷贝。在分析的20例血液系统肿瘤患者中,从患有肺泡蛋白沉积症的白血病患者、2例移植物抗宿主病(GvHD)患者、1例CMV间质性肺炎患者和1例霍奇金病患者获得的两份标本中,CMV病毒DNA水平较高。我们的结果表明,使用实时PCR测量BAL细胞中的CMV基因组不仅有助于了解CMV在系统性呼吸道疾病中的作用,也有助于血液系统肿瘤患者呼吸并发症的护理管理。

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