2017-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656868摘要：乳腺炎是產後最常見的併發症，約可發生於5-33%的產後婦女。但最新的研究發現乳腺炎也可以發生於非孕期、非授乳時期。而有些研究指出此類乳腺炎可能與自體免疫疾病有關，也有研究認為與corynebacterium 感染有關。傳統上corynebacterium 被視為皮膚體表菌，因此一般在皮膚軟組織感染並不會提供進一步的抗生素感受性測試結果。但在本研究團隊的先期研究結果中發現，在某醫學中心2012-2015 年的乳腺炎世代研究中，複雜性(須住院治療)乳腺炎病患中若驗出corynebacterium，是統計學上顯著的危險因子可能會導致較長的治療時間，以及2.96 倍的再復發率。因此我們的假說是corynebacterium 可以是非孕期、非授乳時期乳腺炎之致病菌，本計畫將以前瞻性的世代研究，觀察大於二十歲之乳腺炎病患，以In situ DNA hybridization method 在病理學組織檢測是否可於感染部位發現corynebacterium spp. 存在，並以膿液檢體進行細菌培養及抗生素感受性分析，並以16s rRNA sequencing 進行corynebacterium 菌種確認。若能確認corynebacterium 的角色，本研究進一步的假說是corynebacterium 相關乳腺炎治療成效不佳的原因可能是：未接受有效的對抗corynebacterium 抗生素治療、或是有些抗生素種類無法在相對高脂溶性的乳房組織，達到足夠的抗生素濃度。因此我們將擴展先期的回溯性世代研究至2012-2016 年，針對corynebacterium 相關乳腺炎，分析接受corynebacterium 有效的抗生素治療的有無, 是否為影響預後的顯著影響因子。並在前瞻性研究中，以膿液或乳腺組織進行HPLC 層析儀檢測抗生素濃度，與血清中抗生素濃度比較，分析何種抗生素可於乳房組織達到較佳的組織濃度。若此研究計畫能證明corynebacterium 存在於乳腺炎深部組織中，我們將有進一步的證據建議若於乳腺炎中鑑定出corynebacterium，則應進一步提供species 鑑定及抗生素感受性測試結果，並同時參考不同抗生素在乳腺組織穿透效果，提供病患最適合的抗生素治療，以減少乳腺炎病患的住院天數、避免反覆開刀、及預防乳腺炎的復發。<br> Abstract: Mastitis is one of the most common complications seen in postpartum women, with a prevalence rate of5-33%. However, increasing evidence pointed out the mastitis can appeared among the non-gestational andnon-puerperal stage. Granulomatous mastitis (GM) may related to autoimmune process and some studiessuggest some type of GM may be associated with corynebacterium infection.However, corynebacterium was usually regarded as normal flora and the pathogenic role in mastitis isnot clearly defined. From our preliminary cohort study in a tertially hospital, we had noted the patients who hadcorynebacterium identified in mastitis were associated with longer treatment duration and higher recurrencerate.Thus, we hypothesis that the corynebacterium can be true pathogen in mastitis. The reason for poortreatment outcome or recurrence may be related to inappropriate treatment or low antibiotics concentration inthe lipotropic breast tissue. Through a prospective cohort, we aim to enroll non-gestational and non-puerperalmastitis patients. In situ DNA hybridization method will be used to identify possible corynebacterium infectionin mastitis tissue. Culture of the pus and 16S rRNA gene sequencing will be used to detect and confirmation ofthe corynebacterium. Possible virulence factors among corynebacterium spp., including SpaC, PknG, andNanH will be detected in aspirated pus sample by PCR. The tissue and pus from the infection site will bebiopsy/aspirated to measure the local antibiotics concentration by HPLC method, compared with serumantibiotics level.We also aim to extend the prior retrospective mastitis cohort and focus on corynebacterium associatedmastitis and compare the treatment outcome between the appropriatedly treated group (corynebacteriumsusceptible antibiotics) and non-appropriatedly treated group.Through the study, we can provide more evidence whether corynebacterium can be the true pathogen formastitis. If the study result support our hypothesis, we will further suggest that the corynebacterium should beidentified to the species level and provide antimicrobial susceptibility test result to facilitate the treatment,reduce repeated debridement, and prevent recurrent mastitis.乳腺炎是產後最常見的併發症，約可發生於5-33%的產後婦女。但最新的研究發現乳腺炎也可以 發生於非孕期、非授乳時期。而有些研究指出此類乳腺炎可能與自體免疫疾病有關，也有研究認為與 corynebacterium 感染有關。 傳統上corynebacterium 被視為皮膚體表菌，因此一般在皮膚軟組織感染並不會提供進一步的抗 生素感受性測試結果。但在本研究團隊的先期研究結果中發現，在某醫學中心2012-2015 年的乳腺炎 世代研究中，複雜性(須住院治療)乳腺炎病患中若驗出corynebacterium，是統計學上顯著的危險因 子可能會導致較長的治療時間，以及2.96 倍的再復發率。 因此我們的假說是corynebacterium 可以是非孕期、非授乳時期乳腺炎之致病菌，本計畫將以前 瞻性的世代研究，觀察大於二十歲之乳腺炎病患，以In situ DNA hybridization method 在病理學 組織檢測是否可於感染部位發現corynebacterium spp. 存在，並以膿液檢體進行細菌培養及抗生素 感受性分析，並以16s rRNA sequencing 進行corynebacterium 菌種確認。 若能確認corynebacterium 的角色，本研究進一步的假說是corynebacterium 相關乳腺炎治療成 效不佳的原因可能是：未接受有效的對抗corynebacterium 抗生素治療、或是有些抗生素種類無法在 相對高脂溶性的乳房組織，達到足夠的抗生素濃度。因此我們將擴展先期的回溯性世代研究至 2012-2016 年，針對corynebacterium 相關乳腺炎，分析接受corynebacterium 有效的抗生素治療的 有無, 是否為影響預後的顯著影響因子。並在前瞻性研究中，以膿液或乳腺組織進行HPLC 層析儀檢 測抗生素濃度，與血清中抗生素濃度比較，分析何種抗生素可於乳房組織達到較佳的組織濃度。 若此研究計畫能證明corynebacterium 存在於乳腺炎深部組織中，我們將有進一步的證據建議若 於乳腺炎中鑑定出corynebacterium，則應進一步提供species 鑑定及抗生素感受性測試結果，並同 時參考不同抗生素在乳腺組織穿透效果，提供病患最適合的抗生素治療，以減少乳腺炎病患的住院天 數、避免反覆開刀、及預防乳腺炎的復發。Mastitis is one of the most common complications seen in postpartum women, with a prevalence rate of 5-33%. However, increasing evidence pointed out the mastitis can appeared among the non-gestational and non-puerperal stage. Granulomatous mastitis (GM) may related to autoimmune process and some studies suggest some type of GM may be associated with corynebacterium infection. However, corynebacterium was usually regarded as normal flora and the pathogenic role in mastitis is not clearly defined. From our preliminary cohort study in a tertially hospital, we had noted the patients who had corynebacterium identified in mastitis were associated with longer treatment duration and higher recurrence rate. Thus, we hypothesis that the corynebacterium can be true pathogen in mastitis. The reason for poor treatment outcome or recurrence may be related to inappropriate treatment or low antibiotics concentration in the lipotropic breast tissue. Through a prospective cohort, we aim to enroll non-gestational and non-puerperal mastitis patients. In situ DNA hybridization method will be used to identify possible corynebacterium infection in mastitis tissue. Culture of the pus and 16S rRNA gene sequencing will be used to detect and confirmation of the corynebacterium. Possible virulence factors among corynebacterium spp., including SpaC, PknG, and NanH will be detected in aspirated pus sample by PCR. The tissue and pus from the infection site will be biopsy/aspirated to measure the local antibiotics concentration by HPLC method, compared with serum antibiotics level. We also aim to extend the prior retrospective mastitis cohort and focus on corynebacterium associated mastitis and compare the treatment outcome between the appropriatedly treated group (corynebacterium susceptible antibiotics) and non-appropriatedly treated group. Through the study, we can provide more evidence whether corynebacterium can be the true pathogen for mastitis. If the study result support our hypothesis, we will further suggest that the corynebacterium should be identified to the species level and provide antimicrobial susceptibility test result to facilitate the treatment, reduce repeated debridement, and prevent recurrent mastitis.乳腺炎原位雜合(In Situ HybridizationISH)治療性藥物濃度檢驗棒狀桿菌屬 (Corynebacterium)mastitiscorynebacteriumdrug monitoring