Quantitative determination by real-time PCR of four vaginal Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae indicates an inverse relationship between L. gasseri and L. iners
Quantitative determination by real-time PCR of four vaginal Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae indicates an inverse relationship between L. gasseri and L. iners
复制标题
DOI：
10.1186/1471-2180-7-115
复制
发表时间：
2007-12-19
期刊：
影响因子：
4.2
通讯作者：
Vaneechoutte, Mario
中科院分区：
文献类型：
--
作者：
De Backer, Ellen;Verhelst, Rita;Vaneechoutte, Mario
背景：大多数关于阴道菌群的研究都是基于培养或定性分子技术。本研究采用现有的crispatus乳杆菌、L. gasseri和阴道加德纳菌实时荧光定量PCR格式，并开发了新的阴道托泊菌、L. iners和L. jensen的荧光定量PCR格式，对年龄在18 - 45岁之间的32名孕妇和28名非孕妇的71份阴道样本进行了非培养的定量测定。结果：使用Verhelst等人（2005）改进的Ison和Hay标准，对这些妇女的71例阴道菌群样本进行分类如下：la级：8例，lab级：10例，lb级：13例，I-like级：10例，II级：11例，III级：12例，IV级：7例。除5份样品外，其余样品均检出crispatus，是检出最多的乳杆菌种类。II级和III级的crispatus浓度均显著低于i级（p < 0.0001）和III级（p = 0.002），但所有等级的实验室中均有jensenii浓度高于la级（p = 0.024）。III级时阴道芽孢杆菌和阴道芽孢杆菌浓度较高，中位浓度（log(10) MC）分别为9.0和9.2个细胞/ml。52份阴道支原体阳性标本中有20份（38.5%）阴道支原体阳性。在II级中，我们几乎没有发现L. iners (log(10) MC: 0/ml)，但高浓度的L. gasseri （log(10) MC: 8.7/ml）。相比之下，在III级，我们发现高浓度的L. iners （log(10) MC: 8.3/ml）和低浓度的L. gasseri （log(10) MC: 0/ml）。结果表明，乳杆菌与乳杆菌呈负相关（r = -0.397, p = 0.001），与阴道乳杆菌呈负相关（r = -0.408, p < 0.0001）。结论：本研究发现，乳杆菌与乳杆菌呈明显负相关，阴道乳杆菌与乳杆菌呈明显负相关。我们的研究结果并不支持一般认为的二级是一级和三级之间的中间阶段的命题，因为二级丰富的L. gasseri在三级样品中并不占优势，而三级丰富的L. iners在二级样品中只少量存在。
Background: Most studies of the vaginal microflora have been based on culture or on qualitative molecular techniques. Here we applied existing real-time PCR formats for Lactobacillus crispatus, L. gasseri and Gardnerella vaginalis and developed new formats for Atopobium vaginae, L. iners and L. jensenii to obtain a quantitative non culture-based determination of these species in 71 vaginal samples from 32 pregnant and 28 non-pregnant women aged between 18 and 45 years.Results: The 71 vaginal microflora samples of these women were categorized, using the Ison and Hay criteria, as refined by Verhelst et al. (2005), as follows: grade la: 8 samples, grade lab: 10, grade lb: 13, grade I-like: 10, grade II: 11, grade III: 12 and grade IV: 7. L. crispatus was found in all but 5 samples and was the most frequent Lactobacillus species detected. A significantly lower concentration of L. crispatus was found in grades II (p < 0.0001) and III (p = 0.002) compared to grade I. L. jensenii was found in all grades but showed higher concentration in grade lab than in grade la (p = 0.024). A. vaginae and G. vaginalis were present in high concentrations in grade III, with log(10) median concentrations (log(10) MC), respectively of 9.0 and 9.2 cells/ml. Twenty (38.5%) of the 52 G. vaginalis positive samples were also positive for A. vaginae. In grade II we found almost no L. iners (log(10) MC: 0/ml) but a high concentration of L. gasseri (log(10) MC: 8.7/ml). By contrast, in grade III we found a high concentration of L. iners (log(10) MC: 8.3/ml) and a low concentration of L. gasseri (log(10) MC: 0/ml). These results show a negative association between L. gasseri and L. iners (r = -0.397, p = 0.001) and between L. gasseri and A. vaginae (r = -0.408, p < 0.0001).Conclusion: In our study we found a clear negative association between L. iners and L. gasseri and between A. vaginae and L. gasseri. Our results do not provide support for the generally held proposition that grade II is an intermediate stage between grades I and III, because L. gasseri, abundant in grade II is not predominant in grade III, whereas L. iners, abundant in grade III is present only in low numbers in grade II samples.