Rapid and economical detection of eight carbapenem-resistance genes in Enterobacteriaceae, Pseudomonas spp, and Acinetobacter spp directly from positive blood cultures using an internally controlled multiplex-PCR assay

Multidrug resistant Staphylococcus aureus strains are a major health care problem both in humans and animals. In this work we described three multiplex PCR assays for detection of clinically relevant antibiotic resistance genes in S. aureus isolated from dog dental plaques. Thirty dental plaque samples were collected; then cultural, biochemical and molecular tests performed for isolation and identification of S. aureus from samples. The antibiotic susceptibility of the isolates were checked by Kirby Bauer disc diffusion method and the prevalence of antibiotic resistance genes determined using multiplex PCR assay. As a result S. aureus was isolated from 18 dog plaque samples. Fifteen of these isolates were resistant to penicillin. The mecA gene was more prevalent than blaZ among penicillin-resistant bacteria. Ten of the isolates were resistant to tetracycline. The percentage of tetM was higher than tetK among them. Also, 10 of the isolates were resistant to cefazolin among them bla TEM detected in higher rate than blaSHV and blaOXA-1. Hence multiplex PCR assay is a suitable method for detection of antibiotic resistance patterns of S. aureus isolates.

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Predominance of Acinetobacter spp., Harboring the blaIMP Gene, Contaminating the Hospital Environment in a Tertiary Hospital in Mwanza, Tanzania: A Cross-Sectional Laboratory-Based Study

Pathogens ◽

10.3390/pathogens11010063 ◽

2022 ◽

Vol 11 (1) ◽

pp. 63

Author(s):

Vitus Silago ◽

Eveline C. Mruma ◽

Betrand Msemwa ◽

Conjester I. Mtemisika ◽

Shukurani Phillip ◽

...

Keyword(s):

Carbapenem Resistance ◽

Tertiary Hospital ◽

Hospital Environment ◽

Pcr Assay ◽

Cross Sectional ◽

Middle Income ◽

Carbapenem Resistant ◽

Multiplex Pcr Assay ◽

Genes Encoding ◽

Acinetobacter Spp

Data on colonization and hospital contamination of carbapenem-resistant Gram-negative bacteria (CR-GNB) are limited in low- and middle-income countries. We designed this study to determine the prevalence and co-existence of carbapenemase genes among CR-GNB isolated from clinical, colonization, and hospital environmental samples at a tertiary hospital in Mwanza, Tanzania. The modified Hodge test (MHT), the combined disk test (CDT), and the double-disk synergy test (DDST) were used for the phenotypic detection of carbapenemases. A multiplex PCR assay was used to detect blaIMP and blaKPC, and a singleplex PCR assay was used to detect blaOXA-48. Data were analyzed by STATA version 13.0. Overall, 68.8% (44/64) of the CR-GNB had at least one phenotype by phenotypic methods, whereby 60.9% (39/64) were both CDT and DDST positive and 31.3% (20/64) were MHT positive. A total of 23/64 (35.9%) had at least one of the genes tested with the predominance of blaIMP (91.3%; 21/23). In addition, 47.7% (21/44) of the CR-GNB phenotypes had at least one gene. Around 47.8% (11/23) of the CR-GNB carried multiple genes encoding for carbapenem resistance, with the maximum co-existence of blaIMP/blaKPC/blaOXA-48 (45.5%; 5/11). The majority of carbapenem-resistant genes were detected in Acinetobacter spp. (82.6%; 19/23) and isolated from bed swabs (69.6%; 16/23). Acinetobacter spp. carrying the blaIMP gene predominantly contaminated the hospital environment. Therefore, we recommend routine decontamination of inanimate hospital surfaces, including patient beds.

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A high throughput multiplex PCR assay for simultaneous detection of seven aminoglycoside-resistance genes in Enterobacteriaceae

BMC Microbiology ◽

10.1186/1471-2180-13-58 ◽

2013 ◽

Vol 13 (1) ◽

pp. 58 ◽

Cited By ~ 16

Author(s):

Xiumei Hu ◽

Banglao Xu ◽

Yinmei Yang ◽

Dayu Liu ◽

Mengjie Yang ◽

...

Keyword(s):

Multiplex Pcr ◽

High Throughput ◽

Resistance Genes ◽

Simultaneous Detection ◽

Pcr Assay ◽

Aminoglycoside Resistance ◽

Multiplex Pcr Assay ◽

Aminoglycoside Resistance Genes

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Aetiology of Lobar Pneumonia Determined by Multiplex Molecular Analyses of Lung and Pleural Aspirate Specimens in The Gambia

10.1101/2021.07.02.21259855 ◽

2021 ◽

Author(s):

Grant A Mackenzie ◽

Jessica McLellan ◽

Eunice Machuka ◽

Malick Ndiaye ◽

Jayani Pathirana ◽

...

Keyword(s):

Multiplex Pcr ◽

Pneumococcal Pneumonia ◽

Blood Cultures ◽

Haemophilus Influenzae Type ◽

Haemophilus Influenzae Type B ◽

Type B ◽

Pcr Assay ◽

Lobar Pneumonia ◽

Multiplex Pcr Assay ◽

Vaccine Type

Background Pneumonia aetiology generally relies on insensitive blood cultures or an assumption that organisms in the pharynx are causal. We determined the causes of lobar pneumonia in rural Gambia using lung aspiration. Methods Pneumonia surveillance was undertaken among all ages. Blood culture and chest radiographs were performed routinely while lung or pleural aspirates were collected from selected patients. 7-valent pneumococcal conjugate vaccine (PCV7) was introduced in August 2009 and replaced by PCV13 from May 2011. We used conventional microbiology, and from April 8, 2011 to July 17, 2012, utilized a multiplex PCR assay on lung aspirates. We calculated proportions with pathogens, associations between co-infecting pathogens, and PCV effectiveness. Results 2,550 patients were admitted with clinical pneumonia; 741 with lobar pneumonia or pleural effusion. We performed multiplex PCR on 156 lung and 4 pleural aspirates. Pathogens were detected in 116 specimens, Streptococcus pneumoniae (n=68), Staphylococcus aureus (n=26), and Haemophilus influenzae type b (n=11). Bacteria (n=97) were more common than viruses (n=49). Common viruses were bocavirus (n=11) and influenza (n=11). Co-infections were frequent (n=55). M. catarrhalis was detected in eight patients and in every case there was co-infection with S. pneumoniae. The odds ratio of vaccine-type pneumococcal pneumonia in patients with two or three compared to zero doses of PCV was 0.17 (95% CI 0.06, 0.51). Conclusions Lobar pneumonia in rural Gambia was caused primarily by bacteria, particularly S. pneumoniae and S. aureus. Co-infection was common and M. catarrhalis always co-infected with S. pneumoniae. PCV was highly efficacious against vaccine-type pneumococcal pneumonia.

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Rapid Detection of Bloodstream Pathogens in Oncologic Patients with a FilmArray Multiplex PCR Assay: a Comparison with Culture Methods

Polish Journal of Microbiology ◽

10.5604/01.3001.0011.6149 ◽

2018 ◽

Vol 67 (1) ◽

pp. 103-107 ◽

Cited By ~ 1

Author(s):

Maria Szymankiewicz ◽

Beata Nakonowska

Keyword(s):

Blood Culture ◽

Multiplex Pcr ◽

Accurate Method ◽

Rapid Identification ◽

Blood Cultures ◽

Pcr Assay ◽

Culture Methods ◽

Multiplex Pcr Assay ◽

Oncologic Patients ◽

Culture Identification

The results of the FilmArray® Blood Culture Identification Panel (BCID) (BioFire Diagnostics) and the culture with susceptibility testing of 70 positive blood cultures from oncologic patients were compared. The multiplex PCR assay (BCID) identified 81 of the 83 isolates (97.6%), covered by the panel. The panel produced results in significantly shorter time than standard identification methods, when counted from receiving positive blood cultures bottles to the final results. It is an accurate method for the rapid identification of pathogens and resistance genes from blood culture in oncologic patients.

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Multiplex PCR Assay for Simultaneous Detection of Nine Clinically Relevant Antibiotic Resistance Genes in Staphylococcus aureus

Journal of Clinical Microbiology ◽

10.1128/jcm.41.9.4089-4094.2003 ◽

2003 ◽

Vol 41 (9) ◽

pp. 4089-4094 ◽

Cited By ~ 277

Author(s):

B. Strommenger ◽

C. Kettlitz ◽

G. Werner ◽

W. Witte

Keyword(s):

Staphylococcus Aureus ◽

Antibiotic Resistance ◽

Multiplex Pcr ◽

Resistance Genes ◽

Simultaneous Detection ◽

Antibiotic Resistance Genes ◽

Pcr Assay ◽

Multiplex Pcr Assay

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Rapid Identification of Penicillin and Macrolide Resistance Genes and Simultaneous Quantification of Streptococcus pneumoniae in Purulent Sputum Samples by Use of a Novel Real-Time Multiplex PCR Assay

Journal of Clinical Microbiology ◽

10.1128/jcm.00051-08 ◽

2008 ◽

Vol 46 (7) ◽

pp. 2384-2388 ◽

Cited By ~ 16

Author(s):

K. Y. Fukushima ◽

K. Yanagihara ◽

Y. Hirakata ◽

K. Sugahara ◽

Y. Morinaga ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Multiplex Pcr ◽

Real Time ◽

Resistance Genes ◽

Rapid Identification ◽

Macrolide Resistance ◽

Pcr Assay ◽

Simultaneous Quantification ◽

Multiplex Pcr Assay

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Novel Multiplex PCR Assay for Detection of Chlorhexidine-Quaternary Ammonium, Mupirocin, and Methicillin Resistance Genes, with Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci

Journal of Clinical Microbiology ◽

10.1128/jcm.02488-16 ◽

2017 ◽

Vol 55 (6) ◽

pp. 1857-1864 ◽

Cited By ~ 8

Author(s):

Jo-Ann McClure ◽

Johanna Zaal DeLongchamp ◽

John M. Conly ◽

Kunyan Zhang

Keyword(s):

Staphylococcus Aureus ◽

Multiplex Pcr ◽

Resistance Genes ◽

Quaternary Ammonium ◽

Methicillin Resistance ◽

Rrna Gene ◽

Pcr Assay ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Multiplex Pcr Assay

ABSTRACTMethicillin-resistantStaphylococcus aureus(MRSA) is a clinically significant pathogen that is resistant to a wide variety of antibiotics and responsible for a large number of nosocomial infections worldwide. The Agency for Healthcare Research and Quality and the Centers for Disease Control and Prevention recently recommended the adoption of universal mupirocin-chlorhexidine decolonization of all admitted intensive care unit patients rather than MRSA screening with targeted treatments, which raises a serious concern about the selection of resistance to mupirocin and chlorhexidine in strains of staphylococci. Thus, a simple, rapid, and reliable approach is paramount in monitoring the prevalence of resistance to these agents. We developed a simple multiplex PCR assay capable of screeningStaphylococcusisolates for the presence of antiseptic resistance genes for chlorhexidine and quaternary ammonium compounds, as well as mupirocin and methicillin resistance genes, while simultaneously discriminatingS. aureusfrom coagulase-negative staphylococci (CoNS). The assay incorporates 7 PCR targets, including theStaphylococcus16S rRNA gene (specifically detectingStaphylococcusspp.),nuc(distinguishingS. aureusfrom CoNS),mecA(distinguishing MRSA from methicillin-susceptibleS. aureus),mupAandmupB(identifying high-level mupirocin resistance), andqacandsmr(identifying chlorhexidine and quaternary ammonium resistance). Our assay demonstrated 100% sensitivity, specificity, and accuracy in a total of 23 variant antiseptic- and/or antibiotic-resistant control strains. Further validation of our assay using 378 randomly selected and previously well-characterized local clinical isolates confirmed its feasibility and practicality. This may prove to be a useful tool for multidrug-resistantStaphylococcusmonitoring in clinical laboratories, particularly in the wake of increased chlorhexidine and mupirocin treatments.

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