Lo Shyh-Ching, Hung Guo-Chiuan, Li Bingjie, Lei Haiyan, Li Tianwei, Nagamine Kenjiro, Tsai Shien, Zucker Mark J, Olesnicky Ludmilla
Tissue Microbiology Laboratory, Division of Cellular and Gene Therapies, Office of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration Silver Spring, Maryland 20993, USA.
Department of Medicine, Newark Beth Israel Medical Center Newark, New Jersey 07112, USA.
Int J Clin Exp Pathol. 2015 Nov 1;8(11):13834-52. eCollection 2015.
We examined the microbial composition in the diseased lung and early-phase microbial cultures from the blood of a patient with a rapidly progressing fatal pulmonary illness. Although no microbes could be isolated from such cultures during the initial study, the HTS-microbiome study revealed the presence of a unique mixture of alphaproteobacteria, composed mainly of different families of Rhizobiales microbes. Microbial 16S rDNA sequences matching closely to Afipia cberi were identified mainly in the patient's diseased lung tissue, but only rarely in the early-phase blood cultures. Conversely, the high abundance of sequences found in early-phase blood cultures of different broth media matched closely with those of the families Methylobacteriaceae, Phyllobacteriaceae and Sphingomonadaceae. The two species that successfully adapted to grow in a laboratory culture system were A. cberi and Mesorhizobium hominis, which eventually were isolated from a previously cryopreserved blood culture of SP4 broth. Many other species, including members of the Bradyrhizobiaceae and Phyllobacteriaceae families, and all members of the Methylobacteriaceae and Sphingomonadaceae families identified by HTS remained non-cultivated. We developed specific PCR primers and FISH probes, which detected the target Rhizobiales microbes in former blood cultures and autopsy lung tissues. It is unclear what role these Rhizobiales microbes might have played in the patient's complex disease process. However, the above mentioned assays should help in rapidly detecting and identifying these previously unrecognized Rhizobiales microbes in patients.
我们研究了一名患有快速进展性致命肺部疾病患者的患病肺部微生物组成以及早期血液微生物培养情况。尽管在初始研究期间此类培养物中未分离出微生物，但高通量测序微生物组研究揭示了存在一种独特的α-变形菌混合物，主要由不同科的根瘤菌目微生物组成。与阿菲彼亚菌（Afipia cberi）紧密匹配的微生物16S rDNA序列主要在患者的患病肺组织中被鉴定出来，但在早期血液培养物中很少见。相反，在不同肉汤培养基的早期血液培养物中发现的高丰度序列与甲基杆菌科、叶杆菌科和鞘脂单胞菌科的序列紧密匹配。在实验室培养系统中成功适应生长的两个物种是阿菲彼亚菌和人中华根瘤菌（Mesorhizobium hominis），它们最终从之前冷冻保存的SP4肉汤血液培养物中分离出来。许多其他物种，包括慢生根瘤菌科和叶杆菌科的成员，以及通过高通量测序鉴定的甲基杆菌科和鞘脂单胞菌科的所有成员仍未培养出来。我们开发了特异性PCR引物和荧光原位杂交探针，可在前血液培养物和尸检肺组织中检测目标根瘤菌目微生物。目前尚不清楚这些根瘤菌目微生物在患者复杂的疾病过程中可能发挥了什么作用。然而，上述检测方法应有助于快速检测和鉴定患者中这些以前未被识别的根瘤菌目微生物。
