Login Register Cart 0
Generic placeholder image

Recent Patents on Biotechnology

Editor-in-Chief

ISSN (Print): 1872-2083
ISSN (Online): 2212-4012

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Diagnostic Value of Multiplex Polymerase Chain Reaction in Detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from Sepsis in Pediatrics

Author(s): Sara Galeb, Maysaa El Sayed Zaki*, Raghdaa Shrief, Rasha Hassan and Mohamed Rizk

Volume 15, Issue 3, 2021

Published on: 18 July, 2021

Page: [195 - 203] Pages: 9

DOI: 10.2174/1872208315666210719104623

Price: $65

Abstract

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure.

Objectives: The aim of the present study was to evaluate the multiplex polymerase chain reaction (PCR) for detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children with hospital- acquired sepsis compared to the conventional biochemical reactions for identification of these organisms.

Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR with primers specific to the 3 tested bacteria.

Results: Multiplex PCR was positive in 96% of isolates and 4 isolates had negative results. Falsepositive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa and 27 isolates of Stenotrophomonas maltophilia. The diagnostic value of the multiplex PCR compared to the biochemical identification revealed sensitivity 96.04%, specificity 96.9%, positive predictive value 97.00%, negative predictive value 96.00% and accuracy 96.50%.

Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific and accurate. The accuracy differs according to the organisms with 100% accuracy for Acinetobacter baumannii.

Keywords: Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, PCR, sepsis, bacteria.

« Previous Next »
Graphical Abstract

[1]
Singer M, Deutschman CS, Seymour CW, et al. The third international consensus definitions for sepsis and septic shock (Sepsis-3). JAMA 2016; 315(8): 801-10.
[http://dx.doi.org/10.1001/jama.2016.0287] [PMID: 26903338]
[2]
Page DB, Donnelly JP, Wang HE. Community, healthcare, and hospital acquired severe sepsis hospitalizations in the University Health System Consortium. Crit Care Med 2015; 43(9): 1945-51.
[http://dx.doi.org/10.1097/CCM.0000000000001164] [PMID: 26110490]
[3]
Hotchkiss RS, Moldawer LL, Opal SM, Reinhart K, Turnbull IR, Vincent J-L. Sepsis and septic shock. Nat Rev Dis Primers 2016; 2: 16045.
[http://dx.doi.org/10.1038/nrdp.2016.45] [PMID: 28117397]
[4]
Doi Y. Treatment options for carbapenem-resistant Gram-negative bacterial infections. Clin Infect Dis 2019; 69(Suppl. 7): S565-75.
[http://dx.doi.org/10.1093/cid/ciz830] [PMID: 31724043]
[5]
Gilligan PH, Lum G, VanDamme PAR, Whittier S. Burkholderia, stenotrophomonas, ralstonia, brevundimonas, comamonas, delftia, pandoraea, and acidivorax. In: Carroll KC, Pfaller MA, Landry ML, et al, eds. Manual of clinical microbiology. 8th. Washington, DC: ASM Press 2003; I: pp. 729-48.
[6]
Hotta G, Matsumura Y, Kato K, et al. Risk factors and outcomes of Stenotrophomonas maltophilia bacteraemia: a comparison with bacteraemia caused by Pseudomonas aeruginosa and Acinetobacter species. PLoS One 2014; 9(11): e112208.
[http://dx.doi.org/10.1371/journal.pone.0112208] [PMID: 25375244]
[7]
Alqahtani JM. Emergence of Stenotrophomonas maltophilia nosocomial isolates in a Saudi children’s hospital. Risk factors and clinical characteristics. Saudi Med J 2017; 38(5): 521-7.
[http://dx.doi.org/10.15537/smj.2017.5.16375] [PMID: 28439603]
[8]
Minkwitz A, Berg G. Comparison of antifungal activities and 16S ribosomal DNA sequences of clinical and environmental isolates of Stenotrophomonas maltophilia. J Clin Microbiol 2001; 39(1): 139-45.
[http://dx.doi.org/10.1128/JCM.39.1.139-145.2001] [PMID: 11136762]
[9]
Weiss SL, Fitzgerald JC, Balamuth F, et al. Delayed antimicrobial therapy increases mortality and organ dysfunction duration in pediatric sepsis. Crit Care Med 2014; 42(11): 2409-17.
[http://dx.doi.org/10.1097/CCM.0000000000000509] [PMID: 25148597]
[10]
Marco F. Molecular methods for septicemia diagnosis. Enferm Infecc Microbiol Clin 2017; 35(9): 586-92.
[http://dx.doi.org/10.1016/j.eimc.2017.03.002] [PMID: 28427796]
[11]
Altun O, Almuhayawi M, Ullberg M, Özenci V. Clinical evaluation of the FilmArray blood culture identification panel in identification of bacteria and yeasts from positive blood culture bottles. J Clin Microbiol 2013; 51(12): 4130-6.
[http://dx.doi.org/10.1128/JCM.01835-13] [PMID: 24088863]
[12]
Vila J, Gómez MD, Salavert M, Bosch J. Methods of rapid diagnosis in clinical microbiology: Clinical needs. Enferm Infecc Microbiol Clin 2017; 35(1): 41-6.
[http://dx.doi.org/10.1016/j.eimc.2016.11.004] [PMID: 27993417]
[13]
Escudero D, Forcelledo L, Leoz B, et al. Utility of multiplex PCR (FilmArray Blood Culture Identification) in other biological liquids. Detection of Streptococcus pyogenes in brain abscess and synovial fluid. Rev Esp Quimioter 2019; 32(2): 194-7.
[PMID: 30841692]
[14]
Emami A, Pirbonyeh N, Javanmardi F. Molecular rapid diagnostic method for nosocomial non-fermenting Gram-negative bacilli. Am J Clin Microbiol Antimicrob 2020; 3(1): 1047.
[15]
Boyles TH, Wasserman S. Diagnosis of bacterial infection. S Afr Med J 2015; 105(5): 419.
[http://dx.doi.org/10.7196/SAMJ.9647]
[16]
Novosad SA, Sapiano MRP, Grigg C, et al. Vital signs: epidemiology of sepsis: prevalence of health care factors and opportunities for prevention. MMWR Morb Mortal Wkly Rep 2016; 65(33): 864-9.
[http://dx.doi.org/10.15585/mmwr.mm6533e1] [PMID: 27559759]
[17]
Spilker T, Coenye T, Vandamme P, LiPuma JJ. PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. J Clin Microbiol 2004; 42(5): 2074-9.
[http://dx.doi.org/10.1128/JCM.42.5.2074-2079.2004] [PMID: 15131172]
[18]
Rutledge-Taylor K, Matlow A, Gravel D, et al. A point prevalence survey of health care-associated infections in Canadian pediatric inpatients. Am J Infect Control 2012; 40(6): 491-6.
[http://dx.doi.org/10.1016/j.ajic.2011.08.008] [PMID: 22078941]
[19]
Rudan I, Boschi-Pinto C, Biloglav Z, Mulholland K, Campbell H. Epidemiology and etiology of childhood pneumonia. Bull World Health Organ 2008; 86(5): 408-16.
[http://dx.doi.org/10.2471/BLT.07.048769] [PMID: 18545744]
[20]
Hu J, Robinson JL. Systematic review of invasive Acinetobacter infections in children. Can J Infect Dis Med Microbiol 2010; 21(2): 83-8.
[http://dx.doi.org/10.1155/2010/690715] [PMID: 21629616]
[21]
Büyükcam A, Bıçakcıgil A, Cengiz AB, Sancak B, Ceyhan M, Kara A. Stenotrophomonas maltophilia bacteremia in children - A 10-year analysis. Arch Argent Pediatr 2020; 118(3): e317-23.
[PMID: 32470274]
[22]
Ramanan P, Bryson AL, Binnicker MJ, Pritt BS, Patel R. Syndromic panel-based testing in clinical microbiology. Clin Microbiol Rev 2017; 31(1): e00024-17.
[PMID: 29142077]
[23]
Bhatti MM, Boonlayangoor S, Beavis KG, Tesic V. Evaluation of filmarray and verigene systems for rapid identification of positive blood cultures. J Clin Microbiol 2014; 52(9): 3433-6.
[http://dx.doi.org/10.1128/JCM.01417-14] [PMID: 25031445]
[24]
Rabaan AA, Saunar JV, Bazzi AM, Raslan WF, Taylor DR, Al-Tawfiq JA. Epidemiology and detection of acinetobacter using conventional culture and in- house developed PCR based methods. J Infect Public Health 2017; 10(1): 124-8.
[http://dx.doi.org/10.1016/j.jiph.2016.04.015] [PMID: 27185274]
[25]
Shen K, Sayeed S, Antalis P, et al. Extensive genomic plasticity in Pseudomonas aeruginosa revealed by identification and distribution studies of novel genes among clinical isolates. Infect Immun 2006; 74(9): 5272-83.
[http://dx.doi.org/10.1128/IAI.00546-06] [PMID: 16926421]
[26]
Anuj SN, Whiley DM, Kidd TJ, et al. Identification of Pseudomonas aeruginosa by a duplex real-time polymerase chain reaction assay targeting the ecfX and the gyrB genes. Diagn Microbiol Infect Dis 2009; 63(2): 127-31.
[http://dx.doi.org/10.1016/j.diagmicrobio.2008.09.018] [PMID: 19026507]
[27]
Qin X, Emerson J, Stapp J, Stapp L, Abe P, Burns JL. Use of real-time PCR with multiple targets to identify Pseudomonas aeruginosa and other nonfermenting gram-negative bacilli from patients with cystic fibrosis. J Clin Microbiol 2003; 41(9): 4312-7.
[http://dx.doi.org/10.1128/JCM.41.9.4312-4317.2003] [PMID: 12958262]
[28]
Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE. Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev 2000; 13(4): 559-70.
[http://dx.doi.org/10.1128/CMR.13.4.559] [PMID: 11023957]
[29]
Iglewski BH. Pseudomonas. Medical microbiology. (4th.), The University of Texas Medical Branch at Galveston. 1996; Available from: Available from: https://www.ncbi.nlm.nih.gov/books/NBK8326
[30]
Monnet D, Freney J. Method for differentiating Klebsiella planticola and Klebsiella terrigena from other Klebsiella species. J Clin Microbiol 1994; 32(4): 1121-2.
[http://dx.doi.org/10.1128/JCM.32.4.1121-1122.1994] [PMID: 8027329]
[31]
Bergogne-Bérézin E. The increasing role of Acinetobacter species as nosocomial pathogens. Curr Infect Dis Rep 2001; 3(5): 440-4.
[http://dx.doi.org/10.1007/BF03160479] [PMID: 11559464]
[32]
Allen DM, Hartman BJ. Acinetobacter Species. In: Mandell GL, Bennett JE, Dolin R, Eds. Principles and practice of infectious diseases. 5th ed. Philadelphia: Churchill Livingstone 2000; pp. 2339-44.
[33]
Adegoke AA, Stenström TA, Okoh AI. Stenotrophomonas maltophilia as an emerging ubiquitous pathogen: looking beyond contemporary antibiotic therapy. Front Microbiol 2017; 8: 2276.
[http://dx.doi.org/10.3389/fmicb.2017.02276] [PMID: 29250041]

Rights & Permissions Print Cite
Article Metrics
8
1
© 2024 Bentham Science Publishers | Privacy Policy