第七章 討論
7.7 結論與建議
採集空氣中 S. aureus 之最佳培養基為非選擇性TSA，在搭配 Andersen 1-STG 下，其採樣效能顯著優於其他四類選擇性 MSA、BPA、CSA 與 CSM 培養基，而 四類選擇性培養基之採樣效能彼此間無統計上顯著差異。考量未來實場空氣採樣 或後續樣本分析時，可能須使用選擇性培養基以避免過多雜菌於培養基上生長而 干擾採樣或分析結果，故本研究參考四類選擇性培養基之鑑定能力相關文獻、成 份研究與本實驗採樣效能評估結果，認為選擇性培養基中採樣效能相對最高之 MSA 相當適用於進行實場環境之 S. aureus 空氣採樣。採樣時間方面，Andersen 1-STG 搭配 TSA 與 MSA 於採樣時間 3 分鐘時具 S. aureus 最佳空氣採樣效能，其 採樣效能顯著優於採樣6、15、30 與 60 分鐘之結果，而採樣時間介於6-60 分鐘時 其採樣效能彼此間則無顯著差異。
若以收集液為介質採集空氣中 S. aureus，其最佳空氣採樣效能之收集液為 Tween 80 mixture，顯著優於 PBS 與 DW 之結果；而 PBS 之採樣效能亦顯著高於 DW。然有鑑於 Tween 80 mixture 於採樣時間延長時採樣效能不降反升，很可能係 因Tween 80 mixture 促進S. aureus活性甚至增生所致，故本研究認為以Tween 80 mixture 採集空氣中S. aureus 恐有濃度失真之虞。反之，具適當採樣效能且不含提 供細菌增生之養分的PBS，應較能反應環境空氣中 S. aureus 之真實濃度值。採樣 時間方面，以搭配PBS 採樣之前提下，BioSampler 與 AGI-30 之採樣效能皆隨採樣 時間延長而下降。
將所有採樣方法進行統計檢定，結果顯示，BioSampler 搭配 Tween 80 mixture 之採樣效能最佳；BioSampler 搭配 PBS 以及 AGI-30 搭配 Tween 80 mixture 次之；
值得注意的是，AGI-30 搭配 PBS 之採樣效能最差，其 R 值顯著低於 Andersen 1-STG 搭配TSA，並略遜於 Andersen 1-STG 搭配 MSA 之 R 值。
進一步應用現有文獻偵測所得之 S. aureus 空氣濃度數據檢視實場採樣方法，
並考量本研究以BioSampler 搭配 PBS 於不同採樣時間下之採樣效能結果，本研究 對 S. aureus 實場空氣採樣策略之建議，依採樣目的分為兩項：
1. 若採樣目的係進行環境空氣中 S. aureus 濃度監測，本研究建議使用BioSampler 搭配PBS 進行前測。於環境空氣中S. aureus濃度較低之環境，如醫療院所採樣時，
建議前測時間為30 與 60 分鐘，後續將收集液濃縮至 0.2mL 進行培養法分析。於
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無特定 S. aureus污染源或未經消毒控制之環境，如一般住家採樣時，建議前測時 間為6-60 分鐘，其中採樣時間為 6-15 分鐘時，可將收集液濃縮至 0.2 mL；若採樣 時間延長為30-60 分鐘時，可將收集液濃縮至0.2 mL、0.5mL 或 1mL 後再進行培 養法分析。另，於畜產養殖場所等空氣中 S. aureus濃度較高之環境採樣時，則建 議前測時間為3-60 分鐘，收集液可濃縮至 1mL 或直接經適當稀釋以培養法分析。
若採樣時間較短，考量樣本代表性可視環境特性增加樣本數。
2. 若採樣目的係以定性而非定量為目的，欲透過空氣採樣確認空氣中是否存在具 可培養性之 S. aureus，此時建議可以BioSampler搭配Tween 80 mixture 進行 30-60 分鐘之空氣採樣，採樣中並定時補充收集液。
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119 N（0~360min） 36 36 36 N（0~360min） 108
Mean 30.11 30.13 30.12 Mean 30.12 SD 0.20 0.11 0.19 SD 0.17 CV（%） 0.67 0.38 0.63 CV（%） 0.57
120
121
122
123
附表 5 Nebulizer 中 S. aureus 菌液濃度（n = 3）
系統運作時間
（Ti）（min）*
Nebulizer中菌液濃度（Csusp（Ti））（cfu/mL）**
Mean SD CV（%）
Relative concentration***
Test 1 Test 2 Test 3 （%）
0-20 待暴露艙中相對濕度穩定
20 1.20E+04 8.80E+03 1.01E+04 1.03E+04 1.62E+03 15.71 100.00 50 8.35E+03 5.82E+03 6.90E+03 7.02E+03 1.27E+03 18.11 68.06 80 6.20E+03 4.48E+03 5.43E+03 5.37E+03 8.64E+02 16.10 52.02 110 3.77E+03 2.49E+03 3.48E+03 3.24E+03 6.72E+02 20.72 41.46 140 2.76E+03 1.75E+03 2.48E+03 2.33E+03 5.21E+02 22.36 22.57 170 1.89E+03 1.07E+03 1.57E+03 1.51E+03 4.13E+02 27.46 14.59 200 1.54E+03 1.01E+03 1.29E+03 1.28E+03 2.65E+02 20.75 12.39 230 1.18E+03 9.38E+02 1.13E+03 1.08E+03 1.28E+02 11.84 10.48 260 1.00E+03 7.53E+02 8.45E+02 8.67E+02 1.26E+02 14.58 8.40 290 6.90E+02 6.30E+02 7.58E+02 6.93E+02 6.38E+01 9.21 6.71 320 6.15E+02 5.03E+02 5.40E+02 5.53E+02 5.73E+01 10.37 5.36 N（20~320 min） 11 11 11 N（20~320min） 33
Mean 3.64E+03 2.57E+03 3.14E+03 Mean 3.11E+03 SD 3.72E+03 2.69E+03 3.10E+03 SD 3.13E+03 CV（%） 102.19 104.86 98.90 CV（%） 100.48
*Ti：開啟系統壓縮空氣使其產生生物氣膠之運作時間
**Csusp(Ti)：於系統運作時間Ti 時 Nebulizer 中之菌液濃度
***以系統運作 20 分鐘時所測得之 Nebulizer 中菌液濃度為基準，其他系統運作時間所測得的 Nebulizer 中菌液濃度與其相除之百分比
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附表 6 暴露艙空氣中 S. aureus 濃度（n = 3）
於系統運作中之採 樣時間點（min）
採樣時間之中間時間 點（Tm）（min）*
暴露艙空氣中細菌濃度（Cair
（Tm））（CFU/m3）**
Mean SD CV（%）
Relative concentration***
（%） Test 1 Test 2 Test 3
0-20 待暴露艙中相對濕度穩定
20-30 於 Nebulizer 中添加 S. aureus 菌液後，等待經氣懸之 S. aureus 進入暴露艙至穩定
30-50 40 1.68E+03 8.58E+02 1.24E+03 1.26E+03 4.13E+02 32.73 100.00 60-80 70 9.66E+02 5.15E+02 8.00E+02 7.60E+02 2.28E+02 29.95 60.31 90-110 100 5.53E+02 3.91E+02 5.30E+02 4.91E+02 8.72E+01 17.75 38.97 120-140 130 4.10E+02 2.47E+02 3.88E+02 3.48E+02 8.82E+01 25.33 21.38 150-170 160 2.88E+02 1.94E+02 2.47E+02 2.43E+02 4.72E+01 19.41 19.29 180-200 190 2.13E+02 1.09E+02 1.42E+02 1.55E+02 5.30E+01 34.31 12.27 210-230 220 1.55E+02 1.23E+02 1.34E+02 1.37E+02 1.64E+01 11.95 10.91 240-260 250 1.45E+02 1.07E+02 1.25E+02 1.25E+02 1.89E+01 15.10 9.95 270-290 280 1.07E+02 7.64E+01 1.15E+02 9.93E+01 2.02E+01 20.39 7.87 300-320 310 7.07E+01 6.08E+01 8.83E+01 7.33E+01 1.39E+01 19.03 5.81
N（40~310 min） 10 10 10 N（40~310 min） 30 Mean 4.59E+02 2.68E+02 3.81E+02 Mean 3.69E+02
SD 5.08E+02 2.54E+02 3.80E+02 SD 3.89E+02 CV（%） 110.65 94.82 99.60 CV（%） 105.25
*Tm：以 Andersen 1-STG 採樣 20 分鐘之中間時間點
**Cair(Tm)：於系統運作時間Tm 之暴露艙空氣中細菌濃度
***以系統運作 30-50 分鐘時 Andersen 1-STG 所得暴露艙中細菌濃度為基準，其他運作時間所得暴露艙中細菌濃度與其相除之百分比
125
附表 7 採樣效能指標（R）（n = 3）
於系統運作中之採 樣時間點（min）
採樣時間之中間時間 點（min）
採樣效能指標（R）*
Mean SD CV（%）
Relative concentration**
Test 1 Test 2 Test 3 （%）
30-50 40 0.17 0.12 0.15 0.14 0.02 16.83 100.00 60-80 70 0.13 0.10 0.13 0.12 0.02 14.89 84.66 90-110 100 0.11 0.11 0.12 0.11 0.00 3.75 79.92 120-140 130 0.13 0.12 0.13 0.12 0.01 5.55 28.68 150-170 160 0.12 0.14 0.12 0.13 0.01 6.67 89.72 180-200 190 0.12 0.10 0.10 0.11 0.01 11.79 76.80 210-230 220 0.11 0.13 0.11 0.12 0.01 7.14 82.12 240-260 250 0.13 0.13 0.13 0.13 0.00 2.54 90.02 270-290 280 0.13 0.11 0.14 0.13 0.02 12.98 88.64 300-320 310 0.11 0.11 0.14 0.12 0.02 13.95 82.14
N（40~310 min） 10 10 10 N（40~310 min） 30 Mean 0.13 0.12 0.13 Mean 0.12
SD 0.02 0.01 0.01 SD 0.01 CV（%） 12.65 9.91 11.24 CV（%） 11.72 N（70~310 min） 9 9 9 N（70~310 min） 27
Mean 0.12 0.12 0.12 Mean 0.12 SD 0.009 0.012 0.012 SD 0.012 CV（%） 7.26 10.52 10.52 CV（%） 9.68
*採樣效能指標（R）：以Andersen 1-STG 於暴露艙中採樣到之細菌濃度（Cair（Tm））與該次採樣前後之Nebulizer 中菌液濃度（Csusp
（Ti））平均之比值。
**以 Andersen 1-STG 於系統運作 30-50 分鐘時採樣所得之採樣效能為基準，其他運作時間採樣所得之採樣效能與其相除之百分比
126
127
附表 9 暴露艙中高濃度 S. aureus 之粒徑（n = 3）*
Aerodynamic
Diameter（μm） Test 1_ Mean of raw counts（n=30）
128
129
附表 11 暴露艙中中濃度 S. aureus 之粒徑（n = 3）*
Aerodynamic
Diameter（μm） Test 1_ Mean of raw counts（n=30）
130
131
132
附表 12 Andersen 1-STG 空氣採樣原始數據（續）
採樣器
設計懸浮菌 液濃度
（CFU/mL）
培養基 採樣時間
（min）
實際懸浮菌液濃 度（CFU/mL）
原始菌 落數
（CFU）
校正後 菌落數
（CFU）
空氣中細菌濃
度（CFU/m3） R Mean SD CV
（%）
Andersen
1-STG 5 × 103 TSA 60 3.87 × 103 353 856.6 5.05 × 102 0.130 0.132 0.01 7.60 3.54 × 103 348 816.1 4.81 × 102 0.136
2.62 × 103 317 629 3.71 × 102 0.142 1.65 × 103 225 330.6 1.95 × 102 0.118
MSA 3.67 × 103 288 509.2 3.00 × 102 0.082 0.083 0.01 12.61 3.24 × 103 273 458.9 2.70 × 102 0.083
2.55 × 103 259 417.1 2.46 × 102 0.096 1.61 × 103 154 194.4 1.14 × 102 0.071
133
134
135
136
137
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