Multiplex PCR Analysis for Rapid Detection of Klebsiella pneumoniae Carbapenem-Resistant (Sequence Type 258 [ST258] and ST11) and Hypervirulent (ST23, ST65, ST86, and ST375) Strains

Abstract

Carbapenem-resistant and hypervirulent strains have emerged recently. These strains are both hypervirulent and multidrug resistant and may also be highly transmissible and able to cause severe infections in both the hospital and the community. Clinical and public health needs require a rapid and comprehensive molecular detection assay to identify and track the spread of these strains and provide timely infection control information. Here, we develop a rapid multiplex PCR assay capable of distinguishing carbapenem-resistant isolates of sequence type 258 (ST258) and ST11, and hypervirulent ST23, ST65/ST375, and ST86 clones, as well as capsular types K1, K2, K locus type 47 (KL47), and KL64, and virulence genes ,, , and The assay demonstrated 100% concordance with 118 previously genotyped isolates and revealed different populations of carbapenem-resistant and hypervirulent strains in two collections in China and the United States. The results showed that carbapenem-resistant and hypervirulent strains are still rare in the United States, whereas in China, ∼50% of carbapenem-resistant strains carry and virulence genes, which are largely associated with the epidemic ST11 strains. Similarly, a high prevalence of hypervirulent strains was found in carbapenem-susceptible isolates in two Chinese hospitals, but these primarily belong to ST23, ST65/ST375, and ST86, which are distinct from the carbapenem-resistant strains. Taken together, our results demonstrated that this PCR assay can be a useful tool for molecular surveillance of carbapenem-resistant and hypervirulent strains.

In vitro Activity of Apramycin Against Carbapenem-Resistant and Hypervirulent Klebsiella pneumoniae Isolates

“…In this study, we defined an hvKp strain based upon demonstration of a positive string test (hypermucoviscosity) and co-harboring the genes rmpA (rmpA or rmpA2) and iutA as previously (Yu et al, 2018a). All 84 isolates were hypermucoviscous (string test >5 mm) and were positive for the rmpA/rmpA2 and iutA by PCR analysis (Yu et al, 2018b). In addition, 40 unique strains of carbapenem resistant non-hypervirulent K. pneumoniae (CRnon-hvKp) isolates, with negative hypermucoviscous phenotype and negative rmpA/rmpA2 and iutA PCR results, were collected from the same three hospitals, and were included to compare with the susceptibility results of CR-hvKp.…”

<p>Characterization of Phenotypic and Genotypic Traits of <em>Klebsiella pneumoniae</em> from Lung Cancer Patients with Respiratory Infection</p>

“…7 of clonal dissemination of the test strains by multiplex polymerase chain reaction (PCR) assay. 11…”

“…Isolated DNA samples were screened using specific primers for the detection of virulence genes. 11,19,20 The forward primers were used for DNA sequencing.…”

<p>Dynamic Epidemiology and Virulence Characteristics of Carbapenem-Resistant <em>Klebsiella pneumoniae</em> in Wenzhou, China from 2003 to 2016</p>

“…[18][19][20][21] They have been shown to be highly invasive, in that they usually cause severe life-threatening infections which are associated with high morbidity and mortality in both hospital and community. 20,22,23 Hypervirulence of K. pneumoniae was associated with hypermucoviscosity and diagnostic virulence genes such as rmpA and magA. 24 The emergence of hypervirulent lineages carrying antimicrobial resistance genes including carbapenemases is now being increasingly reported: with limited treatment measures and high mortality, they are considered a "super-bug".…”