FISH-Flow Cytometry: A High-Throughput Molecular Ecological Analysis of Intestinal Microbiota

抄録

Until recently, ecological analysis of human intestinal microbiota had been conducted with culture-dependent methods. However, the majority of our intestinal bacteria remain not-yet cultured. Many culture-independent, molecular ecological methods have been developed to investigate real compositions of microbiota in the last decade. Among them, fluorescencein situhybridization (FISH) is one of the most widely used methods, though it is time-consuming and requires laborious counting work of the detected bacteria. Development of FISH coupled with flow cytometry (FISH-FCM) has circumvented this problem. Since the development of FISH-FCM, it has been used in several fields of microbiology such as environmental microbiology and clinical microbiology. It is now being applied to more complex biological samples, such as feces. These days, intestinal microbiota analyses with FISH-FCM are being conducted not only at group or genus level, but also at the species level. Recently, we have succeeded in the development of a FISH-FCM method that is applicable to fecal samples containing high amounts of autofluorescent particles. Further development of specific probes for the bacteria of interest will contribute to the illustration of the real picture of our intestinal microbiota