刘金荣, 刘立雍, 赵顺英, 徐慧, 唐晓蕾, 张建中, 赵飞. 咽拭子与支气管肺泡灌洗液荧光定量PCR检测儿童肺炎支原体感染分析[J]. 疾病监测, 2019, 34(5): 389-393. DOI: 10.3784/j.issn.1003–9961.2019.05.005
引用本文: 刘金荣, 刘立雍, 赵顺英, 徐慧, 唐晓蕾, 张建中, 赵飞. 咽拭子与支气管肺泡灌洗液荧光定量PCR检测儿童肺炎支原体感染分析[J]. 疾病监测, 2019, 34(5): 389-393. DOI: 10.3784/j.issn.1003–9961.2019.05.005
Jinrong Liu, Liyong Liu, Shunying Zhao, Hui Xu, Xiaolei Tang, Jianzhong Zhang, Fei Zhao. Mycoplasma pneumoniae detection in throat swabs and bronchoalveolar lavage fluid in children with mycoplasmal pneumonia by real-time PCR and culture[J]. Disease Surveillance, 2019, 34(5): 389-393. DOI: 10.3784/j.issn.1003–9961.2019.05.005
Citation: Jinrong Liu, Liyong Liu, Shunying Zhao, Hui Xu, Xiaolei Tang, Jianzhong Zhang, Fei Zhao. Mycoplasma pneumoniae detection in throat swabs and bronchoalveolar lavage fluid in children with mycoplasmal pneumonia by real-time PCR and culture[J]. Disease Surveillance, 2019, 34(5): 389-393. DOI: 10.3784/j.issn.1003–9961.2019.05.005

咽拭子与支气管肺泡灌洗液荧光定量PCR检测儿童肺炎支原体感染分析

Mycoplasma pneumoniae detection in throat swabs and bronchoalveolar lavage fluid in children with mycoplasmal pneumonia by real-time PCR and culture

  • 摘要:
    目的了解咽拭子标本以及支气管肺泡灌洗液(BALF)标本在肺炎支原体(MP)临床核酸检测、细菌定量以及实验室分离培养中的效果,初步评估咽拭子作为核酸检测及分离培养标本的意义,并评估MP肺炎患儿的同期咽拭子的MP核酸载量能否准确预测出BALF的MP核酸载量。
    方法采集29例MP肺炎住院患儿的同期咽拭子及BALF,利用人源性β-globin内参基因质控所有标本采集及核酸提取质量,合格标本使用MP荧光定量PCR对同一患儿同期咽拭子及BALF标本中MP进行定量分析,计算两种标本的MP核酸载量。 并对所有标本进行MP培养,比较两类标本的MP阳性培养率、细菌污染率。
    结果标本的β-globin内参基因检测阳性率为100%,咽拭子和BALF标本荧光PCR检测阳性率均为86.2%(25/29),其中咽拭子MP核酸定量范围是3.2×102 拷贝/ml ~ 1.8×107 拷贝/ml,BALF为6.8×102 拷贝/ml ~ 5.4×108 拷贝/ml。 两种方法检测结果一致率为93.1%(27/29)。 按照BALF标本MP载量递增排序后,阳性标本的咽拭子载量没有显示出类似的伴随升高趋势。 BALF标本培养阳性率为65.5%(19/29),标本污染率为6.9%(2/29);咽拭子标本的MP培养阳性率为51.7%(15/29),标本污染率为51.7%(15/29)。
    结论咽拭子可以作为MP荧光PCR定性检测标本使用,但其不适合用于MP的定量研究,这说明咽拭子的MP核酸载量与BALF中载量不存在明显的相关性;BALF相比咽拭子标本培养率高、污染率低,更适合MP分离培养研究。

     

    Abstract:
    ObjectiveTo understand the performance of clinical nucleic acid detection, bacteria load detection and laboratory isolation/culture of Mycoplasma pneumoniae (MP) in throat swab and bronchoalveolar lavage fluid (BALF) samples and evaluate the significance of using throat swab in the nucleic acid detection and isolation/culture and whether the nucleic acid load of MP in throat swabs in children with MP pneumonia could predict the nucleic acid load of MP in BALF.
    MethodsTwenty-nine hospitalized children with MP pneumonia were enrolled, and throat swabs and BALF samples were collected from them, simultaneously. The quality of the specimens and nucleic acid extraction were controlled by human β-globin internal reference gene. Qualified specimens were used for quantitative detection of MP with real-time PCR, and the nucleic acid load was calculated. At the same time, culture was performed for all the specimens, and positive culture rate and the bacteria contamination rate of the two kinds of specimens were compared.
    ResultsThe positive rate of β-globin reference gene detection in all the specimens was 100%. The positive rate of real-time PCR detection in throat swabs and BALF samples were all 86.2% (25/29), and the quantitative range of MP nucleic acid was 3.2×102 copies/ml−1.8×107 copies/ml in the throat swabs and 6.8×102 copies/ml–5.4×108 copies/ml in BALF samples. The consistent rate of the detection results of two kinds of specimens was 93.1% (27/29). Ranking the MP load in BALF samples showed an increase trend, but no similar trend was found for positive throat swabs. The positive rate and contamination rate of throat swabs were 51.7% (15/29) and 51.7% (15/29), and these of BALF samples were 65.5% (19/29) and 6.9% (2/29), respectively.
    ConclusionThroat swab can be used as qualitative test specimen for MP real-time PCR, but it is not suitable for quantitative study, indicating that there was no significant correlation in MP load between throat swab and BALF. Compared with throat swab specimen, BALF has a higher positive culture rate and lower contamination rate, which makes it more suitable for MP isolation research.

     

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