Design and development of primers for detection of Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis.

IF 1.7 Q3 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

GMS Hygiene and Infection Control Pub Date : 2025-04-30 eCollection Date: 2025-01-01 DOI:10.3205/dgkh000545

Leila Azimi, Fatemeh Shirkavand, Shahnaz Armin, Fereshteh Karbasian, Hannan Khodaei

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Abstract

Background: The mortality rate of meningitis is still alarmingly high in certain regions across the globe. The objective of this research is to identify the most effective primers for detecting Streptococcus (S.) pneumoniae, Haemophilus (H.) influenzae, and Neisseria (N.) meningitidis using Real-Time PCR technology.

Materials and methods: Two sets of primers were developed for detecting S. pneumoniae, H. influenzae, and N. meningitidis using the Primer Biosoft Allele ID 7.6 application. The study examined the minimum bacterial copy numbers detectable by each primer, as well as their specificity.

Results: CtrA and hpd2 could detect the 400 copy numbers/ml of H. influenzae, and N. meningitidis and LytA2 could detect the 40 copy numbers/ml of S. pneumoniae. The sensitivity and specificity of all primers was 100% (CI: 95%).

Conclusion: Using more sensitive primers to detect the bacterial agent responsible for causing bacterial meningitis increases the chance of identifying the causative bacteria. The primers designed in this study could identify the selected bacteria with at least 10 times more sensitivity than the currently available commercial diagnostic kits in Iran.

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设计和开发用于检测肺炎链球菌、流感嗜血杆菌和脑膜炎奈瑟菌的引物。

背景：在全球某些地区，脑膜炎的死亡率仍然高得惊人。本研究的目的是利用Real-Time PCR技术确定检测肺炎链球菌（S.）、流感嗜血杆菌（H.）和脑膜炎奈瑟菌（N.）的最有效引物。材料与方法：采用Primer Biosoft等位基因ID 7.6软件，建立了两套检测肺炎链球菌、流感嗜血杆菌和脑膜炎奈瑟菌的引物。该研究检查了每种引物可检测到的最小细菌拷贝数，以及它们的特异性。结果：CtrA和hpd2可检出流感嗜血杆菌400拷贝数/ml，脑膜炎奈瑟菌和LytA2可检出肺炎链球菌40拷贝数/ml。所有引物的敏感性和特异性均为100% （CI: 95%）。结论：采用更灵敏的引物检测引起细菌性脑膜炎的细菌因子，可增加病原菌的鉴定机会。本研究设计的引物鉴定所选细菌的灵敏度比伊朗目前可用的商业诊断试剂盒至少高10倍。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

GMS Hygiene and Infection Control PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH-

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审稿时长

10 weeks