Development and evaluation of a duplex real-time multienzyme isothermal rapid amplification assay for simultaneous detection of hypervirulent Klebsiella pneumoniae K1 and K2 serotypes in spiked blood samples

IF 1.8 4区医学 Q3 INFECTIOUS DISEASES

Diagnostic microbiology and infectious disease Pub Date : 2026-04-01 Epub Date: 2026-01-17 DOI:10.1016/j.diagmicrobio.2026.117280

Zhixiong Duan , Shaohong Yu , Honglin Wang , Chunyan Yang , Xuelian Peng , Shan Shi , Jin Li

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引用次数: 0

Abstract

The emergence of hypervirulent Klebsiella pneumoniae (hvKP), particularly the K1 and K2 serotypes, presents a significant public health threat due to their increased virulence and resistance. Rapid and accurate detection of hvKP is crucial for effective clinical management. This study developed and evaluated a duplex real-time multienzyme isothermal rapid amplification (MIRA) assay for the simultaneous detection of hvKP K1 and K2 serotypes in spiked blood samples. The assay employed optimized primers and probes targeting specific capsular genes. Sensitivity and specificity were assessed using spiked blood specimens and compared to real-time PCR. The detection limit was 1 × 10³ CFU per reaction for both serotypes, with no cross-reactivity with non-hvKP K1/K2 strains. The assay demonstrated superior reproducibility and stability, offering faster detection and simplified infrastructure requirements compared to real-time PCR. These features make the duplex real-time MIRA assay a promising tool for clinical diagnostics and outbreak surveillance.

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用于同时检测加标血液样本中高毒肺炎克雷伯菌K1和K2血清型的双实时多酶等温快速扩增方法的建立和评价

高致病性肺炎克雷伯菌（hvKP）的出现，特别是K1和K2血清型，由于其毒性和耐药性增强，构成了重大的公共卫生威胁。快速准确地检测hvKP对于有效的临床管理至关重要。本研究开发并评估了一种双重实时多酶等温快速扩增（MIRA）方法，用于同时检测加标血液样本中的hvKP K1和K2血清型。实验采用优化的引物和探针靶向特定的荚膜基因。灵敏度和特异性采用加标血样进行评估，并与实时PCR进行比较。两种血清型的检出限均为1 × 10³CFU /反应，与非hvkp K1/K2菌株无交叉反应性。与实时PCR相比，该分析具有卓越的重复性和稳定性，提供更快的检测速度和简化的基础设施要求。这些特点使双工实时MIRA测定成为临床诊断和疫情监测的有前途的工具。

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

Diagnostic microbiology and infectious disease 医学-传染病学

CiteScore

5.30

自引率

3.40%

发文量

149

审稿时长

56 days

期刊介绍： Diagnostic Microbiology and Infectious Disease keeps you informed of the latest developments in clinical microbiology and the diagnosis and treatment of infectious diseases. Packed with rigorously peer-reviewed articles and studies in bacteriology, immunology, immunoserology, infectious diseases, mycology, parasitology, and virology, the journal examines new procedures, unusual cases, controversial issues, and important new literature. Diagnostic Microbiology and Infectious Disease distinguished independent editorial board, consisting of experts from many medical specialties, ensures you extensive and authoritative coverage.