葡萄糖非發酵菌(glucose nonfermenters)為嗜氧性革蘭氏陰性桿菌，廣泛存在於各種環境中，部分菌種常造成院內感染及伺機性感染。近年來臨床上葡萄糖非發酵菌之抗藥性有逐年增加的情形，有文獻指出以生化反應鑑定葡萄糖非發酵菌，鑑定率可能低於60%。本研究之目的在發展一寡核苷酸晶片(oligonucleotide array)，期能正確鑑定臨床上重要的葡萄糖非發酵菌。本研究之晶片共鑑定487菌株(127菌種)，包括390目標菌株(38菌種，包含119參考菌株及271臨床菌株)及97株非目標菌株(89種)。晶片鑑定結果有16菌株和原菌種名不相符(3株參考菌株及13株臨床菌株)，這些菌株進一步以ITS及16S rRNA基因序列釐清其菌種。本研究發展的晶片鑑定靈敏度(sensitivity)為100% (384/384)，特異性(specificity)為97.0% (96/99)。本晶片可以正確鑑定臨床上常見之葡萄糖非發酵菌，包括一些難以生化反應鑑定的菌種，例如Acinetobacter calcoaceticus-A. baumannii complex (Acb complex)。另外，本研究鑑定307株自成大醫院分離之鮑氏不動桿菌(Acinetobacter baumannii)血液感染分離株，結果顯示有234株(76.2%)是A. baumannii, 70株(22.8%)是Acinetobacter genomic species 3, 2 株(0.7%)是Acinetobacter genomic species 13TU，其餘1株為Acinetobacter sp.。總言之，此晶片可正確鑑定臨床重要葡萄糖非發酵菌，可做為傳統方法的重要輔助方法。

Glucose nonfermenters are gram-negative bacilli and are commonly found in nature environment. They are important opportunistic and nosocomial pathogens. Resistance of nonfermenters to a variety of antibiotics has been recognized worldwide. Recent studies have demonstrated that the identification rates of glucose nonfermenters by phenotypic tests could be below 60%. To solve this problem, an oligonucleotide array was developed to identify glucose nonfermenters. A total of 487 strains (127 species) including 390 target strains (38 species, including 119 reference strains and 271 clinical isolates) and 97 nontarget strains (89 species) were analyzed by the array. There were 16 strains (3 reference strains and 13 clinical isolates) produced discrepant identification between biochemical tests and array hybridization. The discordant strains were analyzed by sequencing of the ITS and 16S rRNA gene for species clarification. The sensitivity and specificity of the array was 100% (384/384) and 97.0% (96/99), respectively. The chip could accurately identify some nonfermenters that are difficult to be differentiated by phenotypic traits, such as species in the Acinetobacter calcoaceticus-A. baumannii complex (Acb complex). By using the array, 307 bacteremic isolates of Acb complex obtained in National Cheng Kung University Hospital were analyzed. Of these 307 isolates, 234 (76.2%), 70 (22.8%), and 2 (0.7%) isolates were identified to be A. baumannii, Acinetobacter genomic species 3, and Acinetobacter genomic species 13TU, respectively, with the remaining one isolate being a strain of Acinetobacter sp. In conclusion, the array could be used as an accurate alternative of conventional methods for identification of clinical glucose nonfermenters.

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