M. Turkey, A., A. Sulaiman, A. (2017). Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance. , 9(2), 73-80. doi: 10.37652/juaps.2015.127656
Ahmed M. Turkey; Ahmed A. Sulaiman. "Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance". , 9, 2, 2017, 73-80. doi: 10.37652/juaps.2015.127656
M. Turkey, A., A. Sulaiman, A. (2017). 'Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance', , 9(2), pp. 73-80. doi: 10.37652/juaps.2015.127656
M. Turkey, A., A. Sulaiman, A. Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance. , 2017; 9(2): 73-80. doi: 10.37652/juaps.2015.127656

Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance

Journal of University of Anbar for Pure Science
Article 19, Volume 9, Issue 2, 2015, Pages 73-80    PDF (622.36 K)
Document Type: Research Paper
DOI: 10.37652/juaps.2015.127656
Authors
Ahmed M. Turkey* 1; Ahmed A. Sulaiman2
1University of Anbar / College of Science
2University of Anbar/ Center for Desert Studies
Abstract
450 samples were collected from different pathological cases (urine, wounds, Burns, stool, and nasal and pharyngeal swabs) from November 2014 through February 2015. 60 samples were diagnosed as Staph. aureus and 35 samples were Strep. pyogenes using phenotypic, cultural and biochemical diagnosis features and definitely diagnosed with Vaitek test. Results of antibiotic resistance against different antibiotics showed variations in their resistance to these antibiotics, and 10 isolates were selected of each gender on the basis of variation in antibiotic resistance. A number of primers were designed to detect bacteria use distinct gene and also to detects resistance to antibiotics using multiplex PCR. Agarose gel electrophoresis for polymerization reaction showed that all Staph. aureus isolates had femA which might be used as detection marker where all Strep. pyogenes had Tu(tuf), in the same reaction of multiplex PCR ,detection of seven genes related with resistance to different antibiotic groups (Amp, tetR, Tri, Gen, Imp, Qnl and mecA) were done, all Staph. aureus contained Imp,Gen,mecA, tetR,Tri, and Qnl genes while its varied in their contain of Amp gene . Also all Strep. pyogenes had Tri,Gen,Amp and mecA genes while its varied in their contain of Qnl which not detected in all isolates. From this study it's easy to use multiplex PCR to detect pathogenic bacteria along with their antibiotic resistance without need to long routine work.
Keywords
multiplex PCR; Antibiotic resistance; strep; Pyogenes; staph; aureus
References

1-   Pena. I، Picazo. J.J،Rodrigueu- Avail.C  and  Rodrigueu- Avail. I ( 2014) . J. Antimicrob.Agents 43(5) 460- 464.

2-   Chiu SK،Wu TL, Chuang YC, Lin JC،Fung CP،Lu PL ( 2013 ). Nationalsurveil- lance study on carbapenemnon-susceptible Klebsiella pneumoniae in Taiwan: the emergenceandrapiddisseminationofKPC-2carbapenemase.PLoSOne  ;8:e69428.

3-   Bexiga, R., Koskinen, M.T., Holopainen, J., Carneiro, C., Pereira, H., Ellis, K.A.,Vilela, C.L.,( 2011). Diagnosis of intramammary infection in samples yielding negative results or minor pathogens in conventional bacterial culturing. J. Dairy Res. 78, 49–55.

4- التومی، عبد الرزاق سلیمان، محمد  محمد الامام، عبد الباسط رمضان ابو زویده  (2013). اساسیات التشخیص البکتیریولوجی المعملی والسریری، مرکز بحوث التقنیات.

5-   Stefani S, Chung DR and Lindsay JA. (2012).Methicillin resistant Staphylococcus aureus (MRSA) :Global Epidemiology and Harmonisation of typing methods.International ajaournal of Antimicrobial Agent.39(4):273-282.

6-   Chiara Montanari ، Diana I. Serrazanetti ، Giovanna Felis ، Sandra Torriani ، Giulia Tabanelli ، Rosalba Lanciotti  and Fausto Gardini. (2015 ).  New insights in thermal resistance of staphylococcal strains belonging to the species Staphylococcus epidermidis, Staphylococcus lugdunensis and Staphylococcus aureus. J Food Control ; 50 : 605 – 612.

7-   Hwang SY,Kim sh, Jang EJ,Kwon NH,Park YK K HC،Jung wk،kim J.M. and parkY.H.(2007).Novel multiplex PCR for detection of Staphylococcus aureus superantigen and its application to raw meat isolates in korea.J.food Microb. 3941:1_7.

8-   Al-Talib.  Hassanain، Chan Yean Yean، Alyaa Al-Khateeb، Habsah Hasan, Manickam Ravichandran (2014). Rapid detection of methicillin-resistant Staphylococcus aureus by a newly developed dry reagent-based polymerase chain reaction assay. Journal of Microbiology, Immunology and Infection. 47, 484- 490.

9-   Podbielksi and Andreas.(2007). Flexible architecture of streptococcus pyogenes FeT Genome region : Finally the clue for under standing purulent skin disease and long term persistence., J Backeriol. 189 : 1181 – 1184.

10-    Iciar Rodriguez-Avial, Carmen Rodriguez-Avial ، Esther Culebras, Juan J. Picazo, (2003). Distribution of tetracycline resistance genes tet(M), tet(O), tet(L) and tet(K) in blood isolates of viridans group streptococci harbouring erm(B) and mef (A) genes. Susceptibility to quinupristin /dalfopristin and linezolid. International Journal of Antimicrobial Agents ; 21 : 536-541.

11-    Danbing.Ke، Maurice Biossinot، Ann Huletsky، Francois Picard، Johanne Frentte، Marc oullette، Paul H.Ray and Michel G. Bergeron (2000). Evidence for horizontal gene transfer in evolution of elongation factor Tu in enterococci. J Bacterial.  182 (24) : 6913 -6920.

12-    Baron،E.J. and Finedgold، S.M. (1990) Diagnostic microbiology. 8th ed. Mosby –Year – Book. Inc. Missouri. USA.

13-    Holt, J.G.; Krieg, N.R.; Sneath, P.H.A.; Stalyt, J.T. and Williams, S.T. (1994). Bergey`s manual of determinative bacteriology. Williams and Wilkins Publication. 9th ed. London, New York.

14-    Vandepitte, J. ; Engback, K. ; Piot, P. and Heuck, C.C.(1991).Basic laboratory procedures in clinical bacteriology. WHO., Geneva.، PP: 78-110.

15-    Arif,Sehand K. and Salih, layla I.F. (2010). Identification of Different    Categories of Diarrheagenic Escherichia coli in Stool Samples by Using Multiplex PCR Technique. J. Medical Sciences. 2(5): 237-243.

16-    Blahna MT, Zalewski CA, Reuer J, Kahlmeter G, Foxman B, Marrs CF ( 2006 ). The role of horizontal gene transfer in the spread of trimethoprim–sulfamethoxazole resistance among uropathogenic Escherichia coli in Europe and Canada. J Antimicrob Chemother ;57:666–72.

17-    Melican, K. ; Sandoval, R. M. ; Abdul- Kader ; Josefsson, L. ; Tanner, G. A. ; Molitoris, B. A. and Richter-Dahlfors, A. (2011). Uropathogenic Escherichia coli P and type 1 fimbriae act in synergy in a living host to facilitate renal colonization leading to nephron obstruction. PLoS Pathogens، 7 (2): 100-112.

18-    Francis. M, Francois j. p، Louis G ، Paul H. Roy، Marc Ouellette ( 2000) . Multiplex PCR assays for the detection of clinically relevant antibiotic resistance genes in staphylococci isolated from patient sinfected after cardiac surgery. j. of Antimicrobial chemotheraoy ; 46 : 527- 533.

19-    Al-Talib H، Yean CY,  Al -Khteeb A, Hassan H, Singh KB, Al- Jashamy K, Ravichandran A (2009). Apentaplex PCR assay for the rapid detection of methicillin resistant Staphylococcus aureus and Panton _Valentine Leucocidine. J.BMC Microb. 9:1471-2180.

20-    Birgit Strommenger, Christiane Kettlitz, Guido Werner, and Wolfgang Witte., ( 2003). Multiplex PCR Assay for Simultaneous Detection of Nine Clinically Relevant Antibiotic Resistance Genes in Staphylococcus aureus. journal of clinical microbiology; 41 ( 9) : 4089–4094.

21-    Su Mi Choi, Seung-Han Kim،Hee-Jung Kim، Dong-Gun Lee, Jung-Hyun Choi, Jin-Hong Yoo, Jin-Han Kang، Wan-Shik Shin, Moon-Won Kang.( 2003). Multiplex PCR for the Detection of Genes Encoding Aminoglycoside Modifying Enzymes and Methicillin Resistance among Staphylococcus Species. J Korean Med Sci ; 18: 631-6.

22-    Nizami Duran, Burcin Ozer, Gulay Gulbol Duran، Yusuf Onlen and  Cemil Demir (2012).  Antibiotic resistance genes and  susceptibility patterns in staphylococci.  Indian J Med Res. 135 : 389-396.

23-    Surbhi Malhotra-Kumar,  Christine Lammens, Jasper Piessens, and Herman Goossens.(2005). Multiplex PCR for Simultaneous Detection of Macrolide and Tetracycline Resistance Determinants in Streptococci. antimicrobial agents and chemotherapy,  49 (11)، p. 4798–4800.

24-    Louis Davignon,  Elizabeth A. Walter,  Kate M. Mueller,  Christopher. P and Barrozo.(2005). Use of Resequencing Oligonucleotide Microarrays for Identification of Streptococcus pyogenes and Associated Antibiotic Resistance Determinants. journal of clinical microbiology, 43 (11) : 5690–5695.

25-    Vanesa Reijtman، Paula Gagetti، Diego Faccone، Sofía Fossati، Patricia Sommerfleck, Claudia Hernández, Patricia Bernáldez, Horacio Lopardo, and Alejandra Corso. (2013). Macrolide resistance in Streptococcus pneumoniae isolated from Argentinian pediatric patients suffering from acute otitis media. J Rev Argent Microbiol ;45(4):262-266.

26-    Corso A, Faccone D, Galia C, Gagetti P, Rodríguez M, Pace J and Regueira M, (2009). The Argentinean SIREVA working group. Prevalenceof mef and ermB genes in invasive pediatric eritromycinresistance Streptococcus pneumoniae isolates from Argentina. Rev Argent Microbiol.;41:29-33.

 

 

Statistics
Article View: 424
PDF Download: 226


Journal Management System. Powered by iJournalPro.com