Rachana Solanki1, Lavanya Vanjari2, Sreevidya Subramanian3, Aparna B4, Nagapriyanka E5, Vemu Lakshmi6. 1. Senior Resident, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India . 2. Research Scholar, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India . 3. Junoir Resident, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India . 4. Technician, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India . 5. Junior Research Fellow, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India . 6. Professor & Head, Department of Microbiology, Nizam's Institute of Medical Science , Panjagutta, Hyderabad, Telangana, India .
Abstract
BACKGROUND: Carbapenem resistant pathogens cause infections associated with significant morbidity and mortality. OBJECTIVE: This study evaluates the use of Multiplex PCR for rapid detection of carbapenemase genes among carbapenem resistant Gram negative bacteria in comparison with the existing phenotypic methods like modified Hodge test (MHT), combined disc test (CDT) and automated methods. MATERIAL AND METHODS: A total of 100 Carbapenem resistant clinical isolates, [Escherichia coli (25), Klebsiella pneumoniae (35) P. aeruginosa (18) and Acinetobacter baumannii (22)] were screened for the presence of carbapenemases (bla NDM-1, bla VIM , blaIMP and blaKPC genes) by phenotype methods such as the modified Hodge test (MHT) and combined disc test (CDT) and the molecular methods such as Multiplex PCR. RESULTS: Seventy of the 100 isolates were MHT positive while, 65 isolates were positive by CDT. All the CDT positive isolates with EDTA and APB were Metallo betalactamase (MBL) and K. pneumoniae carbapenemase (KPC) producers respectively. bla NDM-1 was present as a lone gene in 44 isolates. In 14 isolates bla NDM-1 gene was present with blaKPC gene, and in one isolate bla NDM-1 gene was present with blaVIM , gene. Only one E. coli isolate had a lone blaKPC gene. We didn't find bla IMP gene in any of the isolates. Neither of the genes could be detected in 35 isolates. CONCLUSION: Accurate detection of the genes related with carbapenemase production by Molecular methods like Multiplex PCR overcome the limitations of the phenotypic methods and Automated systems.
BACKGROUND:Carbapenem resistant pathogens cause infections associated with significant morbidity and mortality. OBJECTIVE: This study evaluates the use of Multiplex PCR for rapid detection of carbapenemase genes among carbapenem resistant Gram negative bacteria in comparison with the existing phenotypic methods like modified Hodge test (MHT), combined disc test (CDT) and automated methods. MATERIAL AND METHODS: A total of 100 Carbapenem resistant clinical isolates, [Escherichia coli (25), Klebsiella pneumoniae (35) P. aeruginosa (18) and Acinetobacter baumannii (22)] were screened for the presence of carbapenemases (bla NDM-1, bla VIM , blaIMP and blaKPC genes) by phenotype methods such as the modified Hodge test (MHT) and combined disc test (CDT) and the molecular methods such as Multiplex PCR. RESULTS: Seventy of the 100 isolates were MHT positive while, 65 isolates were positive by CDT. All the CDT positive isolates with EDTA and APB were Metallo betalactamase (MBL) and K. pneumoniae carbapenemase (KPC) producers respectively. bla NDM-1 was present as a lone gene in 44 isolates. In 14 isolates bla NDM-1 gene was present with blaKPC gene, and in one isolate bla NDM-1 gene was present with blaVIM , gene. Only one E. coli isolate had a lone blaKPC gene. We didn't find bla IMP gene in any of the isolates. Neither of the genes could be detected in 35 isolates. CONCLUSION: Accurate detection of the genes related with carbapenemase production by Molecular methods like Multiplex PCR overcome the limitations of the phenotypic methods and Automated systems.
Authors: Karthikeyan K Kumarasamy; Mark A Toleman; Timothy R Walsh; Jay Bagaria; Fafhana Butt; Ravikumar Balakrishnan; Uma Chaudhary; Michel Doumith; Christian G Giske; Seema Irfan; Padma Krishnan; Anil V Kumar; Sunil Maharjan; Shazad Mushtaq; Tabassum Noorie; David L Paterson; Andrew Pearson; Claire Perry; Rachel Pike; Bhargavi Rao; Ujjwayini Ray; Jayanta B Sarma; Madhu Sharma; Elizabeth Sheridan; Mandayam A Thirunarayan; Jane Turton; Supriya Upadhyay; Marina Warner; William Welfare; David M Livermore; Neil Woodford Journal: Lancet Infect Dis Date: 2010-08-10 Impact factor: 25.071
Authors: George L Daikos; Panayiotis Petrikkos; Mina Psichogiou; Chris Kosmidis; Evangelos Vryonis; Athanasios Skoutelis; Kleoniki Georgousi; Leonidas S Tzouvelekis; Panayotis T Tassios; Christina Bamia; George Petrikkos Journal: Antimicrob Agents Chemother Date: 2009-02-17 Impact factor: 5.191
Authors: K F Anderson; D R Lonsway; J K Rasheed; J Biddle; B Jensen; L K McDougal; R B Carey; A Thompson; S Stocker; B Limbago; J B Patel Journal: J Clin Microbiol Date: 2007-06-20 Impact factor: 5.948
Authors: Fred C Tenover; Rajinder K Kalsi; Portia P Williams; Roberta B Carey; Sheila Stocker; David Lonsway; J Kamile Rasheed; James W Biddle; John E McGowan; Bruce Hanna Journal: Emerg Infect Dis Date: 2006-08 Impact factor: 6.883
Authors: Ana P Silva; Isabel Faria-Ramos; Elisabete Ricardo; Isabel M Miranda; Maria J Espinar; Sofia Costa-de-Oliveira; Rafael Cantón; Acácio G Rodrigues; Cidália Pina-Vaz Journal: Antimicrob Agents Chemother Date: 2016-05-23 Impact factor: 5.191