Comparison of phenotypic and PCR methods for detection of carbapenemases production by Enterobacteriaceae

AlTamimi, Maryam; Al‐Salamah, Ali A.; AlKhulaifi, Manal; AlAjlan, Hisham

doi:10.1016/j.sjbs.2016.07.004

Cited by 36 publications

(36 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The current study revealed the superiority of MHT over Carba NP test for detection of carbapenemases, this finding was also reported by a recent Egyptian study 35 that showed a high sensitivity for MHT in which 100% (14/14) of CRKP isolates, that were molecularly confirmed to be carbapenemase producers, were tested positive for MHT. Similar findings were reported by AlTamimi et al 40 who found that the sensitivity of MHT among CRKP isolates was 92% (23/25). In the current study, we supposed that the negative MHT result for the five isolates that genotypically tested positive for carbapenemases may be due to false negatives rather than true negatives.…”

Section: Discussionsupporting

confidence: 91%

See 1 more Smart Citation

The First Egyptian Report Showing the Co-Existence of blaNDM-25, blaOXA-23, blaOXA-181, and blaGES-1 Among Carbapenem-Resistant K. pneumoniae Clinical Isolates Genotyped by BOX-PCR

El-Badawy¹,

El‐Far²,

Althobaiti³

et al. 2020

IDR

View full text Add to dashboard Cite

Background and Objective: The emergence of carbapenem-resistant K. pneumoniae (CRKP) continues to escalate and is alarming because of the emergence of pan drugresistant strains. The objective of this study was to investigate the existence of 12 carbapenemase genes among CRKP clinical isolates. Methods: Ninety-six Klebsiella spp. clinical isolates were collected. The isolates were identified phenotypically and genotypically. These isolates were screened for susceptibility to 24 different antibiotics. The modified Hodge test (MHT) and the Carba Nordmann/Poirel (NP) test were used to phenotypically screen carbapenem-resistant strains for carbapenemase production. Phenotypic characterization of carbapenemases was performed using the combined disk synergy test (CDST). Additionally, the presence of 12 carbapenemase genes in CRKP isolates was investigated. The DNA sequence of bla NDM and bla GES genes was determined. The BOX-PCR technique was used to determine the clonal relationship between CRKP isolates. Results: All carbapenem-resistant isolates were related to K. pneumoniae. Susceptibility testing showed that 19.79% (19/96) of the collected isolates were carbapenem-resistant. Of the CRKP isolates, 68.42% (13/19) tested positive for the MHT and Carba NP test. CDST showed that 42.11% (8/19), 63.16% (12/19), 47.37% (9/19), and 73.68% (14/19) of the CRKP isolates tested positive for the inhibitory effect of clavulanic acid, sulbactam, phenylboronic acid, and tazobactam, respectively, while 84.21% (16/19) and 68.42% (13/16) tested positive for the inhibitory effect of EDTA and mercaptopropionic acid, respectively. It was found that 10.53% (2/19) of the isolates tested positive for the inhibitory effect of sodium chloride. Molecular investigation of carbapenemases showed that 26.32% (5/19), 73.68% (14/19), 21.05% (4/19), 10.53% (2/19), and 5.26% (1/19) of the isolates tested positive for bla NDM , bla OXA-48 , bla OXA-181 , bla OXA-51 , and bla OXA-23 , respectively. None of the isolates tested positive for bla OXA-40 and bla OXA-58. Two allelic variants of bla NDM (bla NDM-1 and bla NDM-25) were detected. BOX-PCR revealed high clonal relatedness between CRKP isolates. Conclusion: MHT was more sensitive than Carba NP test for evaluating carbapenemase production and class D carbapenemase genes were the most prevalent of the 12 carbapenemase genes that were evaluated.

show abstract

Section: Discussionsupporting

confidence: 91%

“…Of the detected OTCs, OXA-48 was the most prevalent one in which 73.68% (14/19) of isolates were found to harbor bla OXA-48 . Similar findings were reported by the Turkish, 43 Egyptian, 31 and Saudi 40,44 studies that reported that bla OXA-48 was more prevalent in CRKP clinical isolates.…”

Section: Discussionsupporting

confidence: 90%

The First Egyptian Report Showing the Co-Existence of blaNDM-25, blaOXA-23, blaOXA-181, and blaGES-1 Among Carbapenem-Resistant K. pneumoniae Clinical Isolates Genotyped by BOX-PCR

El-Badawy¹,

El‐Far²,

Althobaiti³

et al. 2020

IDR

View full text Add to dashboard Cite

show abstract

“…Recent studies conducted by Al Tamimi et al . and ElMahallawy et al ., had reported (43%) and(49%), respectively with the MHT results [ 27 – 28 ]. In the current study, relatively low percentage of carbapenemase detection by MHT could be attributed to presence of class B carbapenemase including NDMs, VIMs and IMPs that requires zinc for their action[ 22 ].…”

Section: Discussionmentioning

confidence: 94%

Phenotypic screening and molecular characterization of carbapenemase-producing Gram-negative bacilli recovered from febrile neutropenic pediatric cancer patients in Egypt

et al. 2018

View full text Add to dashboard Cite

AimInfections with carbapenem-resistant Gram-negative bacteria (GNB) are among the most frequent complications in the immunocompromised cancer patients because of their considerable morbidity and mortality. Therefore, the aim of the current study was to characterize the prevalence of carbapenemase-producing GNB recovered from febrile neutropenic pediatric cancer patients in Egypt.MethodsStandard methods were used for identification, sensitivity testing (Kirby-Bauer and broth microdilution method according to CLSI guidelines). Standard methods were applied for both phenotypic and genotypic detection of the carbapenemase-producing GNB.ResultsA total of 185 GNB were recovered from different clinical specimens, Escherichia (E.) coli (86; 46.48%), followed by Klebsiella spp. (71; 38.37%), Acinetobacter (A.) baumannii (7; 3.78%) and others including Pseudomonas spp., Enterobacter (Ent.) cloacae and Proteus spp. (21; 11.35%). It is a matter of concern that 116 out of 171 enterobacterial isolates (94.15%) showed resistance to three or more antimicrobial classes and were considered multidrug resistant. Additionally, the rate of carbapenem-resistance displayed a worrisome trend as 113 out of 171 enterobacterial isolates (66.08%) and 12 out of 14 non fermenting bacilli (85.71%) showed resistance pattern to at least one of the tested carbapenems. After performing a series of phenotypic tests for initial screening of potential carbapenemase producers, molecular characterization to the 29 extracted plasmids were subjected to PCR (using 5 common carbapenemase primers). The results revealed that blaOXA-48 was the most prevalent 17 (58.62%), followed by blaNDM 8(27.58%), then blaVIM 3 (10.3%) and blaKPC 2 (6.89%).ConclusionThese results are an alarming threat to public health that calls for urgent application of antimicrobial stewardship programs along with routine surveillance for controlling outbreaks.

show abstract

“…These include Xpert Carba-R (Cepheid), Check-Direct CPE (Check-Points Health), and an antibiotic resistance TaqMan assay (ThermoFisher Scientific). Xpert Carba-R is a fully automated and integrated system for sample preparation, DNA extraction, amplification, and qualitative detection of target genes using multiplex real-time PCR assays with a Ͻ1-h turnaround time (214). The method can be used directly on swab specimens and has 96.6% sensitivity and 98.6% specificity (215).…”

Section: Detection Of Ndm-encoding Genesmentioning

confidence: 99%

NDM Metallo-β-Lactamases and Their Bacterial Producers in Health Care Settings

et al. 2019

View full text Add to dashboard Cite

SUMMARYNew Delhi metallo-β-lactamase (NDM) is a metallo-β-lactamase able to hydrolyze almost all β-lactams. Twenty-four NDM variants have been identified in >60 species of 11 bacterial families, and several variants have enhanced carbapenemase activity.Klebsiella pneumoniaeandEscherichia coliare the predominant carriers ofblaNDM, with certain sequence types (STs) (forK. pneumoniae, ST11, ST14, ST15, or ST147; forE. coli, ST167, ST410, or ST617) being the most prevalent. NDM-positive strains have been identified worldwide, with the highest prevalence in the Indian subcontinent, the Middle East, and the Balkans. MostblaNDM-carrying plasmids belong to limited replicon types (IncX3, IncFII, or IncC). Commonly used phenotypic tests cannot specifically identify NDM. Lateral flow immunoassays specifically detect NDM, and molecular approaches remain the reference methods for detectingblaNDM. Polymyxins combined with other agents remain the mainstream options of antimicrobial treatment. Compounds able to inhibit NDM have been found, but none have been approved for clinical use. Outbreaks caused by NDM-positive strains have been reported worldwide, attributable to sources such as contaminated devices. Evidence-based guidelines on prevention and control of carbapenem-resistant Gram-negative bacteria are available, although none are specific for NDM-positive strains. NDM will remain a severe challenge in health care settings, and more studies on appropriate countermeasures are required.

show abstract

Comparison of phenotypic and PCR methods for detection of carbapenemases production by Enterobacteriaceae

Cited by 36 publications

References 36 publications

<p>The First Egyptian Report Showing the Co-Existence of <em>bla</em><sub>NDM-25</sub>, <em>bla</em><sub>OXA-23</sub>, <em>bla</em><sub>OXA-181</sub>, and <em>bla</em><sub>GES-1</sub> Among Carbapenem-Resistant <em>K. pneumoniae</em> Clinical Isolates Genotyped by BOX-PCR</p>

<p>The First Egyptian Report Showing the Co-Existence of <em>bla</em><sub>NDM-25</sub>, <em>bla</em><sub>OXA-23</sub>, <em>bla</em><sub>OXA-181</sub>, and <em>bla</em><sub>GES-1</sub> Among Carbapenem-Resistant <em>K. pneumoniae</em> Clinical Isolates Genotyped by BOX-PCR</p>

Phenotypic screening and molecular characterization of carbapenemase-producing Gram-negative bacilli recovered from febrile neutropenic pediatric cancer patients in Egypt

NDM Metallo-β-Lactamases and Their Bacterial Producers in Health Care Settings

Contact Info

Product

Resources

About