Abstract: This research examines the poor villages released in the Thirteen-Five Planning of Hubei, Hunan and Jiangxi provinces in the middle reaches of the Yangtze River. The Specific Methods of Nuclear Density Analysis (SMDNA), Nearest Neighbor Index (SNI), Kernel Density (KD), Moran Index (MI) and Overlay Analysis (OA) and other spatial analysis methods were selected to analyze the quantity, spatial distribution density and pattern, and vertical differentiation of the poor villages. The results show that: (1) the inter-provincial distribution of poor villages is different in the middle reaches of the Yangtze River. Hunan having the largest number, followed by Hubei and Jiangxi province, and there are typical “plain” poor villages and “mountainous” poor villages, and the quantitative characteristics are different at multi-level scales such as provincial, national poor counties, provincial poor counties, non-poor counties and urban circles; (2) the distribution of poor villages is low-value clustered in urban groups or urban circles, high-value clustered in mountainous area at the provincial borders; even in the contiguous poor areas of Wushan, Wulingshan and Xuefengshan; and significantly clustered in Jianghan Plain; (3) the vertical space differentiation of poor villages is significant too. The poor villages are mainly distributed in plain area with altitude below 200m, and with the increase of altitude, the number of poor villages is decreasing. The research helps to better understand the spatial pattern of poor villages in the middle reaches of the Yangtze River and implement a more scientific ‘targeted poverty alleviation’ strategy.