Ferris Michael J, Masztal Alicia, Aldridge Kenneth E, Fortenberry J Dennis, Fidel Paul L, Martin David H
Dept of Pediatrics, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
BMC Infect Dis. 2004 Feb 13;4:5. doi: 10.1186/1471-2334-4-5.
Bacterial vaginosis (BV) is a polymicrobial syndrome characterized by a change in vaginal flora away from predominantly Lactobacillus species. The cause of BV is unknown, but the condition has been implicated in diverse medical outcomes. The bacterium Atopobium vaginae has been recognized only recently. It is not readily identified by commercial diagnostic kits. Its clinical significance is unknown but it has recently been isolated from a tuboovarian abcess.
Nucleotide sequencing of PCR amplified 16S rRNA gene segments, that were separated into bands within lanes on polyacrylamide gels by denaturing gradient gel electrophoresis (DGGE), was used to examine bacterial vaginal flora in 46 patients clinically described as having normal (Lactobacillus spp. predominant; Nugent score < or = 3) and abnormal flora (Nugent score > or = 4). These women ranged in age from 14 to 48 and 82% were African American.
The DGGE banding patterns of normal and BV-positive patients were recognizably distinct. Those of normal patients contained 1 to 4 bands that were focused in the centre region of the gel lane, while those of BV positive patients contained bands that were not all focused in the center region of the gel lane. More detailed analysis of patterns revealed that bands identified as Atopobium vaginae were present in a majority (12/22) of BV positive patients, while corresponding bands were rare (2/24) in normal patients. (P < 0.001) Two A. vaginae isolates were cultivated from two patients whose DGGE analyses indicated the presence of this organism. Two A. vaginae 16S rRNA gene sequences were identified among the clinical isolates. The same two sequences were obtained from DGGE bands of the corresponding vaginal flora. The sequences differed by one nucleotide over the short (approximately 300 bp) segment used for DGGE analysis and migrated to slightly different points in denaturing gradient gels. Both isolates were strict anaerobes and highly metronidazole resistant.
The results suggest that A. vaginae may be an important component of the complex bacterial ecology that constitutes abnormal vaginal flora. This organism could play a role in treatment failure if further studies confirm it is consistently metronidozole resistant.
细菌性阴道病(BV)是一种多微生物综合征,其特征是阴道菌群从以乳酸杆菌为主发生改变。BV的病因尚不清楚,但该病症与多种医学结局有关。阴道阿托波菌直到最近才被认识。它不易被商业诊断试剂盒鉴定。其临床意义尚不清楚,但最近已从输卵管卵巢脓肿中分离出该菌。
通过变性梯度凝胶电泳(DGGE)将PCR扩增的16S rRNA基因片段在聚丙烯酰胺凝胶的泳道内分离成条带,然后进行核苷酸测序,以检查46例临床诊断为阴道菌群正常(以乳酸杆菌属为主; Nugent评分≤3)和异常(Nugent评分≥4)的患者的阴道菌群。这些女性年龄在14至48岁之间,82%为非裔美国人。
正常患者和BV阳性患者的DGGE条带模式明显不同。正常患者的条带模式包含1至4条条带,集中在凝胶泳道的中央区域,而BV阳性患者的条带模式包含的条带并非都集中在凝胶泳道的中央区域。对条带模式的更详细分析显示,在大多数(12/22)BV阳性患者中存在被鉴定为阴道阿托波菌的条带,而在正常患者中相应条带很少见(2/24)。(P<0.001)从两名DGGE分析表明存在该菌的患者中培养出两株阴道阿托波菌。在临床分离株中鉴定出两个阴道阿托波菌16S rRNA基因序列。从相应阴道菌群的DGGE条带中获得了相同的两个序列。在用于DGGE分析的短(约300 bp)片段上,这两个序列相差一个核苷酸,并且在变性梯度凝胶中迁移到略有不同的位置。两株分离株均为严格厌氧菌,对甲硝唑高度耐药。
结果表明,阴道阿托波菌可能是构成异常阴道菌群的复杂细菌生态的重要组成部分。如果进一步研究证实其对甲硝唑持续耐药,该菌可能在治疗失败中起作用。