通过实时PCR对四种阴道乳酸杆菌、阴道加德纳菌和阴道阿托波菌进行定量测定，结果表明加氏乳杆菌和惰性乳杆菌之间呈负相关。
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.
作者信息
De Backer Ellen, Verhelst Rita, Verstraelen Hans, Alqumber Mohammed A, Burton Jeremy P, Tagg John R, Temmerman Marleen, Vaneechoutte Mario
机构信息
Department of Clinical Chemistry, Microbiology & Immunology, Ghent University Hospital, UGent, Ghent, Belgium.
出版信息
BMC Microbiol. 2007 Dec 19;7:115. doi: 10.1186/1471-2180-7-115.
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 Ia: 8 samples, grade Iab: 10, grade Ib: 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 Iab than in grade Ia (p = 0.024). A. vaginae and G. vaginalis were present in high concentrations in grade III, with log10 median concentrations (log10 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 (log10 MC: 0/ml) but a high concentration of L. gasseri (log10 MC: 8.7/ml). By contrast, in grade III we found a high concentration of L. iners (log10 MC: 8.3/ml) and a low concentration of L. gasseri (log10 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.
背景
大多数关于阴道微生物群的研究都是基于培养或定性分子技术。在此，我们应用现有的针对卷曲乳杆菌、加氏乳杆菌和阴道加德纳菌的实时聚合酶链反应（PCR）方法，并开发了针对阴道阿托波菌、惰性乳杆菌和詹氏乳杆菌的新方法，以对来自32名年龄在18至45岁之间的孕妇和28名非孕妇的71份阴道样本中的这些菌种进行基于非培养的定量测定。
结果
根据Verhelst等人（2005年）改进的Ison和Hay标准，将这些女性的71份阴道微生物群样本分类如下：Ia级：8份样本，Iab级：10份，Ib级：13份，I类：10份，II级：11份，III级：12份，IV级：7份。除5份样本外，其余样本中均发现了卷曲乳杆菌，且它是检测到的最常见的乳杆菌种类。与I级相比，II级（p < 0.0001）和III级（p = 0.002）中卷曲乳杆菌的浓度显著降低。在所有级别中均发现了詹氏乳杆菌，但Iab级中的浓度高于Ia级（p = 0.024）。阴道阿托波菌和阴道加德纳菌在III级中浓度较高，对数中位数浓度（log10 MC）分别为9.0和9.2个细胞/毫升。在52份阴道加德纳菌阳性样本中，有20份（38.5%）也对阴道阿托波菌呈阳性。在II级中，我们几乎未发现惰性乳杆菌（log10 MC：0/毫升），但加氏乳杆菌浓度较高（log10 MC：8.7/毫升）。相比之下，在III级中，我们发现惰性乳杆菌浓度较高（log10 MC：8.3/毫升），而加氏乳杆菌浓度较低（log10 MC：0/毫升）。这些结果表明加氏乳杆菌与惰性乳杆菌之间（r = -0.397，p = 0.001）以及加氏乳杆菌与阴道阿托波菌之间（r = -0.408，p < 0.0001）呈负相关。
结论
在我们的研究中，我们发现惰性乳杆菌与加氏乳杆菌之间以及阴道阿托波菌与加氏乳杆菌之间存在明显的负相关。我们的结果不支持普遍认为的II级是I级和III级之间的中间阶段这一观点，因为在II级中大量存在的加氏乳杆菌在III级中并非占主导地位，而在III级中大量存在的惰性乳杆菌在II级样本中仅少量存在。
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