应用套式聚合酶链反应检测支气管肺泡灌洗液中的滴虫。
Use of nested PCR for the detection of trichomonads in bronchoalveolar lavage fluid.
机构信息
The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China.
Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China.
出版信息
BMC Infect Dis. 2019 Jun 10;19(1):512. doi: 10.1186/s12879-019-4118-9.
BACKGROUND
The methods routinely used to detect trichomonads in the lungs are not sensitive enough, and an effective method is urgently needed.
METHOD
Primers were first designed to match the conserved area of the 18S rRNA gene of trichomonads. Then, nested PCR was carried out to detect trichomonads in bronchoalveolar lavage fluid (BALF). Finally, all positive specimens were subjected to DNA sequencing and phylogenetic analysis.
RESULTS
Among 115 bronchoalveolar lavage fluid samples, ten samples tested positive in nested PCR (10/115), while no samples were positive in wet mount microscopy (0/115) (P < 0.01). Among the ten positive specimens, two were identified as Tetratrichomonas spp. and the other eight as Trichomonas tenax in phylogenetic analysis.
CONCLUSIONS
Nested PCR is an effective way to detect trichomonads in bronchoalveolar lavage fluid.
背景
目前用于检测肺部滴虫的方法不够敏感，因此急需一种有效的方法。
方法
首先设计针对滴虫 18S rRNA 基因保守区的引物，然后进行巢式 PCR 检测支气管肺泡灌洗液（BALF）中的滴虫。最后，对所有阳性标本进行 DNA 测序和系统发育分析。
结果
在 115 份支气管肺泡灌洗液样本中，巢式 PCR 检测出 10 份阳性（10/115），而湿片镜检均为阴性（0/115）（P<0.01）。在 10 份阳性标本中，2 份鉴定为四毛滴虫属，其余 8 份鉴定为阴道毛滴虫在系统发育分析中。
结论
巢式 PCR 是检测支气管肺泡灌洗液中滴虫的有效方法。
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