The Center of Clinical Laboratory, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Chin Med J (Engl). 2009 Nov 20;122(22):2748-51.
The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women. Thus an accurate understanding of the composition and ecology of the ecosystem is important to understanding the etiology of these diseases. This study aimed to compare the characteristics of main lactobacillus species between healthy women and women with bacterial vaginosis (BV) by quantitative culture and PCR methods. Main lactobacillus species include L. crispatus, L. gasseri, L. jensenii and L. iners.
A total of 150 women attending Gynecology Outpatient Clinic of Beijing Friendship Hospital, were diagnosed as having BV because three or more of the following criteria were met (standard of Amsel's composite criteria): homogeneous discharge, elevated vaginal pH (pH > 4.5), production of amines, and presence of "clue" cells. Those with less than three of the criteria were considered as healthy. Simultaneously, smears were made of vaginal fluid and Gram stained, then were assessed qualitatively as normal (grade I), intermediate (grade II), or consistent with BV (grade III). Gardnerella vaginalis were identified by using Vitek 2 Compact and PCR methods. Lactobacillus species were identified by PCR methods. Gardnerella vaginalis and lactobacilli colony counts were determined by calculating the most number of colonies of each species in the appropriate plates (colonies between 10 and 300), corrected by the dilution of the sample in the plates, and multiplied by 10 (to account for plating 100 microl), in order to get colony forming units per milliliter of vaginal secretion.
BV was diagnosed in 31% (46/150) patients using the composite criteria, the remainder being regarded as healthy. The majority of patients with BV had a smear assessed as grade III (91%, 42/46) and minority of them had a smear assessed as grade II (9%, 4/46). The majority of healthy women had a smear assessed as grade I (64%, 67/104). Smears assessed as grade II were found (36%, 37/104) among patients diagnosed as healthy, with less than three of the composite criteria. L. crispatus was cultured from 94% of healthy women and 83% of women with BV, with the former colonies count average value of 10(6) and the latter of 10(3). L. gasseri, L. iners, and L. jensenii were cultured from 85%, 68% and 43% of healthy women; and 28%, 89% and 44% of BV women, respectively.
The quantities of four lactobacillus species except L. jensenii had a significant difference between healthy women and women with BV. Our results provide support for the negative association between L. iners and L. gasseri. Although L. crispatus were existent both in healthy and BV positive women's vagina, the numbers of L. crispatus were significantly different for the dominant number in healthy women. Smears of vaginal fluid and Gram stain play an important guiding role in bacteria culture.
阴道正常微生物菌群在预防女性生殖道和尿路感染方面发挥着重要作用。因此，准确了解生态系统的组成和生态学对于了解这些疾病的病因至关重要。本研究旨在通过定量培养和 PCR 方法比较健康女性和细菌性阴道病（BV）女性主要乳杆菌属的特征。主要乳杆菌属包括 L. crispatus、L. gasseri、L. jensenii 和 L. iners。
共有 150 名在北京友谊医院妇科门诊就诊的女性被诊断为 BV，因为满足以下三个或更多标准（Amsel 综合标准）：均质分泌物、阴道 pH 值升高（pH>4.5）、产生胺类物质和存在“线索”细胞。那些少于三个标准的被认为是健康的。同时，对阴道液进行涂片并革兰氏染色，然后定性评估为正常（I 级）、中间（II 级）或符合 BV（III 级）。通过使用 Vitek 2 Compact 和 PCR 方法鉴定加德纳菌。通过 PCR 方法鉴定乳杆菌属。通过计算每个物种在适当平板上的最多菌落数（10 到 300 个菌落之间）来确定加德纳菌和乳杆菌的菌落计数，通过平板中的样品稀释进行校正，并乘以 10（以考虑接种 100 微升），以获得每毫升阴道分泌物中的菌落形成单位。
使用综合标准诊断 BV 患者占 31%（46/150），其余为健康患者。大多数 BV 患者的涂片评估为 III 级（91%，42/46），少数为 II 级（9%，4/46）。大多数健康女性的涂片评估为 I 级（64%，67/104）。在被诊断为健康的患者中发现了涂片评估为 II 级（36%，37/104），他们的综合标准少于三个。L. crispatus 从 94%的健康女性和 83%的 BV 女性中培养出来，前者的菌落计数平均值为 10（6），后者为 10（3）。L. gasseri、L. iners 和 L. jensenii 从 85%、68%和 43%的健康女性中培养出来，从 28%、89%和 44%的 BV 女性中培养出来。
除 L. jensenii 外，四种乳杆菌属的数量在健康女性和 BV 女性之间存在显著差异。我们的结果支持 L. iners 和 L. gasseri 之间的负相关。虽然 L. crispatus 存在于健康和 BV 阳性女性的阴道中，但 L. crispatus 的数量在健康女性中明显不同，是主要菌群。阴道液涂片和革兰氏染色在细菌培养中起着重要的指导作用。
