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題名 | Use Universal Bacterial Primers for Polymerase Chain Reaction to Dectect the Common Pathogens in Neonatal Sepsis=利用聚合酶連鎖反應偵測引起新生兒敗血症的常見細菌 |
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作者 | 侯世婷; 王志堅; 朱夢麟; | 書刊名 | 中華民國感染症醫學會雜誌 |
卷期 | 8:1 1997.06[民86.06] |
頁次 | 頁6-10 |
分類號 | 417.517 |
關鍵詞 | 聚合酶連鎖反應; 偵測; 新生兒; 敗血症; 細菌; Neonatal sepsis; Polymerase chain reaction; 16S rDNA gene; |
語文 | 英文(English) |
中文摘要 | 雖然醫藥的進步,新生兒敗血症仍然是造成其罹病以及死亡的主要因素。由於臨床表徵不典型,診斷新生兒敗血症並不容易;本篇研究,我們嚐試用聚合�t連鎖反應來偵測引起新生兒敗血症的十種常見細菌,包括group B Streptococcus, Staphylococcus aureus, Staphylococcus epidermidis, Steptococcus pneumoniae, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Haemophilus influenzae 以及 Listeria monocytogenes 等。先定量並依序稀釋之後,用煮沸法抽取細菌DNA,所用的引子是:p8FPL和p806R,反應周期如下:先95℃五分鐘;再30周期的95℃一分鐘,55℃一分鐘,以及72 ℃二分鐘;最後在72 ℃十分鐘。 藉放大高度保存於所有細菌內的16S rDNA 基因,找出可被偵測的最低細菌濃度,所得結果,約10至10�揪熊葍q可以被聚合�t連鎖反應偵測到。藉由本篇初步的研究,可以了解放大高度保存於所有細菌基因內16S rDNA的聚合�t連鎖反應之敏感度是非常高的。藉由這些經驗及技術,可以應用到臨床檢體,相信對於早期診斷新生兒敗血症是很有助益的。 |
英文摘要 | Despite improvements in medicine, bacterial sepsis still remains a major cause of morbidity and mortality in newborn infants. For atypical clinical presentation, diagnosis of bacterial infection may be particular difficult in such young patients. In this study, we tried to find the lowest detectable bacterial concentration of common pathogens of neonatal sepsis, including group B Streptococcus, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoriae, Enterococcus feacalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Haemophilus influenzae and Literia monocytogenes, by using the polymerase chain reaction (PCR). The two primers of highly conserved 16S rDNA gene were p8FPL and p806R. Routine amplification conditions were: an initial denaturation for 5 min at 95 ℃; 30 cycles of denaturation at 95 ℃ for 1 min, annealing at 55 ℃ for 1 min, and extension at 72 ℃ for 2 min; and a final extension at 72 ℃. The results were 10 to 10�� bacteria could be detected which made the PCR results positive. This preliminary study give us the sensitivity of PCR for amplification of highly conserved DNA sequences found in all bacteria. It would be an improvement to develop a fast, sensitive and specific method to determine the presence of bacteria in clinical specimens and thus, make it possible to diagnose neonatal sepsis earlier. |
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