Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms.
Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms.
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DOI：
10.1111/j.1574-6968.2010.02100.x
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发表时间：
2010-11
影响因子：
2.1
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Liu G;Saxena D;Deng H;Norman RG;Chen Z;Abrams WR;Malamud D;Li Y
蛋白酶抑制剂混合物通常被添加到用于蛋白质组学研究的临床样品中以抑制蛋白酶。由于这些相同的样品通常用于微生物研究，因此我们确定添加蛋白酶抑制剂是否会影响微生物谱的定量或定性评估。收集了22份唾液样本，并在加入或不加入蛋白酶抑制剂混合物的情况下立即进行处理。常规培养方法用于评价总细菌生长。提取总基因组DNA，PCR扩增特异性16 S rRNA基因靶向区域，变性梯度凝胶电泳分离。结合1D SDS-PAGE和LC-MS/MS方法来确定蛋白酶抑制剂对唾液蛋白和肽的完整性的影响。有趣的是，在两组之间的细菌生长和组成或唾液蛋白的完整性方面没有观察到显著差异。配对样本中总可培养菌群（r2=0.847）、总变形链球菌（r2=0.898）、总口腔乳酸杆菌（r2=0.933）和总变形链球菌（r2=0.870）的相关系数也超过了预期值。结果表明，在唾液样本中添加蛋白酶抑制剂混合物不会影响口腔微生物群的生长或损害表征其组成的能力。
Protease inhibitor cocktails are routinely added to clinical samples used for proteomic studies to inactivate proteases. Since these same samples are often used for microbial studies, we determined whether addition of protease inhibitors could affect the quantitative or qualitative assessment of microbial profiles. Twenty-two saliva samples were collected and processed immediately with or without the addition of a protease inhibitor cocktail. Conventional cultivation methods were used to evaluate total bacterial growth. Total genomic DNA was isolated and a specific 16S rRNA gene-targeted region was PCR-amplified and separated by denaturing gradient gel electrophoresis. A combination of 1D SDS-PAGE and LC-MS/MS methods was used to determine the effect of the protease inhibitors on the integrity of salivary proteins and peptides. Interestingly, no significant differences were observed in either the bacterial growth and composition or the integrity of salivary proteins between the two groups. Correlation coefficients between the paired samples for total cultivable microbiota (r2=0.847), total mutans streptococci (r2=0.898), total oral lactobacilli (r2=0.933), and total Streptococcus mutans (r2=0.870) also exceeded expected values. The results suggest that the addition of a protease inhibitor cocktail in saliva samples does not impact the growth of oral microbiota or compromise the ability to characterize its composition.
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影响因子：
3.1
作者：
Hiemstra, PS;Maassen, RJ;Dijkman, JH
通讯作者：
Dijkman, JH
影响因子：
2.1
作者：
GRENIER, D;MICHAUD, J
通讯作者：
MICHAUD, J
DOI：
10.1111/j.1365-2672.1992.tb01821.x
发表时间：
1992-03-01
期刊：
JOURNAL OF APPLIED BACTERIOLOGY
影响因子：
--
作者：
PELLEGRINI, A;THOMAS, U;WILD, P
通讯作者：
WILD, P
影响因子：
7.6
作者：
Li, Y;Ku, CYS;Caufield, PW
通讯作者：
Caufield, PW
影响因子：
3.1
作者：
Curtis, MA;Opoku, JA;Samuelsson, B
通讯作者：
Samuelsson, B