PCR for Screening of Multidrug Resistant Acinetobacter baumannii by MexA, MexB Like Gene and 16-23S rRNA Intergenic Spacer Region (ISR)
Abstract
This research was aimed to identify MexA and MexB like gene in Acinetobacter baumannii from clinical isolates and compare it among some opportunistic bacteria. The MexA and MexB gene was analyzed and compared by PCR and confirmed the difference of A. baumannii among other species by 16S - 23S rRNA Intergenic Spacer Region (ISR). The results of research could amplify both the MexA and MexB like DNAs from multidrug-resistant A. baumannii of which was not found in nondrug-resistant A. baumannii among all clinical isolates from Rayong Hospitals and Chonburi Hospital, collected during March to December 2016. Thus, the research was revealed that these Mex A and MexB like genes dispread in the two provinces, and these genes contained PCR products of 1,175 and 2,300 base pairs, respectively. Also, identification of A. baumannii by using ISR primer could indicate that twenty isolates of both multidrug-resistant A. baumannii and non multidrug-resistant A. baumannii contained 1,258 base pairs of ISR. Thus, PCR could be applied to select out multidrug-resistant A. baumannii and discrete from other opportunistic bacteria. This method also included to study evolution of resistant bacteria and incidence of outbreaks for treatment and prevention of A. baumannii infection in the future. Keywords : multidrug resistant ; Acinetobacter baumannii ; MexA and MexB like gene ; 16S – 23S rRNA Intergenic Spacer Region (ISR)References
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Blair, J. M., & Piddock, L.J. (2009). Structure, function and inhibition of RND efflux pumps in Gram- negative bacteria: an update, Current Opinion in Microbiology, 12 (5), 512–519.
CLSL. (2017). Performance standards for antibacterial susceptibility testing. 27th ed. CLSL supplements M100. Wayne, PA: Clinical and Laboratory Standards Institute.
Hurley, J.C. (2016). World-wide variation in incidence of Acinetobacter associated ventilator associated pneumonia: a meta-regression. BMC Infectious Diseases, 16, 577, (DOI 10.1186/s12879-016-1921-4).
Ko, W.C., Lee, N.Y., Su, S.C., Dijkshoorn, L., Vaneechoutte, M., Wang, L. R., Yan, J. J., & Chang, T. C. (2008). Oligonucleotide array-base identification of species in the Acinetobacter calcoaceticos-A. baumannii complex in Isolates from blood culture and antimicrobial susceptibility testing of the Isolates. Journal of Clinical Microbiology, 46 (6), 2052-2059.
Lee, S.Y., Shin, J.H., Park, K.H., Kim, J.H., Shin, M.G., Suh, S.P., & Ryang, D.W. (2014). Identification, genotypic relation, and clinical features of colistin-resistant isolatates of Acinetobacter genomic species 13BJ/14TU from bloodstreams of patients in a university hospital. Journal of Clinical Microbiology, 52(3), 931-939.
Lee, C.R. Lee, J.H., Park, M., Park, K.S. , Bae, K., Kim, Y.B., Cha, C.K., Jeong, B.C., & Lee , S.H. (2017). Biology of Acinetobacter baumannii: pathogenesis, antibiotic resistance mechanisms, and prospective treatment options. Frontiers in Cellular and Infection Microbiology, 7, 1-35.
Li, X. Z., Plesiat, P. & Nikaido, H. (2015). The challenge of efflux-mediated antibiotic resistance in gram-negative bacteria. Clinical Microbiology Reviews, 28(2), 337-418.
Lister, D. P., Wolter J. D., & Hanson D. N. (2009). Antibacterial-Resistant Pseudomonas aeruginosa: Clinical Impact and Complex Regulation of Chromosomally Encoded Resistance Mechanisms. Clinical Microbiology reviews, 22 (4), 528-610.
Liu, C., Chang, Y., Xu, Y., Luo, Y., W, L., Mei, Z., Li, S., Wang, R. & Jia, X. (2018). Distribution of virulence- associated genes and antimicrobial susceptibility in clinical Acinetobacter baumannii isolates. Oncotarget, 9(31), 21663-21673.
Minagawa, S., Inami, S., Kato, T., Sawada, S., Yasuki, T., Miyairi, S., Horikawa, M., Okuda, J., & Gotoh, N. (2012). RND type efflux pump system MexAB-OprM of Pseudomonas aeruginosa selects bacterial languages,
3-oxo-acyl-homoserine lactones, for cell-to-cell communication. BMC Microbiology, 12 (70), 1471-2180.
Ogawa, W., Onishi, M., Ni, R., Tsuchiya, T., & Kuroda, T. (2012). Functional study of the novel multidrug efflux pump KexD from Klebsiella pneumoniae. Gene, 498(2), 177–182.
Pagdepanichkit , S., Tribuddharat C., & Chuanchuen R. (2016) Distribution and expression of the Ade multidrug efflux systems in Acinetobacter baumannii clinical isolates. Canadian Journal of Microbiology, 62(9) 794-801.
Puttilerpong, C., Chawanasit, W.,Laohawaleesan, W, Rungsang, W., Ritteeverkul, P. & (2011). Antimicroial use in hospital-acquired pneumonia with multidrug-resistant Acinetobacter baumannii at King Chulalongkorn Memorial Hospital. Thai Pharmaceutical and Health Science Journal, 6(1), 32-38. (in Thai)
Wieczorek, P., Sacha, P.,.Hauschild, T.,Zórawski, M., Krawczyk, M., & Tryniszewska, E. (2008). Multidrug resistant Acinetobacter baumannii-the role of AdeABC (RND family) efflux pump in resistance to antibiotics. Foloa Histichemica et Cytochemica, 46(3), 257-267.
Blair, J. M., & Piddock, L.J. (2009). Structure, function and inhibition of RND efflux pumps in Gram- negative bacteria: an update, Current Opinion in Microbiology, 12 (5), 512–519.
CLSL. (2017). Performance standards for antibacterial susceptibility testing. 27th ed. CLSL supplements M100. Wayne, PA: Clinical and Laboratory Standards Institute.
Hurley, J.C. (2016). World-wide variation in incidence of Acinetobacter associated ventilator associated pneumonia: a meta-regression. BMC Infectious Diseases, 16, 577, (DOI 10.1186/s12879-016-1921-4).
Ko, W.C., Lee, N.Y., Su, S.C., Dijkshoorn, L., Vaneechoutte, M., Wang, L. R., Yan, J. J., & Chang, T. C. (2008). Oligonucleotide array-base identification of species in the Acinetobacter calcoaceticos-A. baumannii complex in Isolates from blood culture and antimicrobial susceptibility testing of the Isolates. Journal of Clinical Microbiology, 46 (6), 2052-2059.
Lee, S.Y., Shin, J.H., Park, K.H., Kim, J.H., Shin, M.G., Suh, S.P., & Ryang, D.W. (2014). Identification, genotypic relation, and clinical features of colistin-resistant isolatates of Acinetobacter genomic species 13BJ/14TU from bloodstreams of patients in a university hospital. Journal of Clinical Microbiology, 52(3), 931-939.
Lee, C.R. Lee, J.H., Park, M., Park, K.S. , Bae, K., Kim, Y.B., Cha, C.K., Jeong, B.C., & Lee , S.H. (2017). Biology of Acinetobacter baumannii: pathogenesis, antibiotic resistance mechanisms, and prospective treatment options. Frontiers in Cellular and Infection Microbiology, 7, 1-35.
Li, X. Z., Plesiat, P. & Nikaido, H. (2015). The challenge of efflux-mediated antibiotic resistance in gram-negative bacteria. Clinical Microbiology Reviews, 28(2), 337-418.
Lister, D. P., Wolter J. D., & Hanson D. N. (2009). Antibacterial-Resistant Pseudomonas aeruginosa: Clinical Impact and Complex Regulation of Chromosomally Encoded Resistance Mechanisms. Clinical Microbiology reviews, 22 (4), 528-610.
Liu, C., Chang, Y., Xu, Y., Luo, Y., W, L., Mei, Z., Li, S., Wang, R. & Jia, X. (2018). Distribution of virulence- associated genes and antimicrobial susceptibility in clinical Acinetobacter baumannii isolates. Oncotarget, 9(31), 21663-21673.
Minagawa, S., Inami, S., Kato, T., Sawada, S., Yasuki, T., Miyairi, S., Horikawa, M., Okuda, J., & Gotoh, N. (2012). RND type efflux pump system MexAB-OprM of Pseudomonas aeruginosa selects bacterial languages,
3-oxo-acyl-homoserine lactones, for cell-to-cell communication. BMC Microbiology, 12 (70), 1471-2180.
Ogawa, W., Onishi, M., Ni, R., Tsuchiya, T., & Kuroda, T. (2012). Functional study of the novel multidrug efflux pump KexD from Klebsiella pneumoniae. Gene, 498(2), 177–182.
Pagdepanichkit , S., Tribuddharat C., & Chuanchuen R. (2016) Distribution and expression of the Ade multidrug efflux systems in Acinetobacter baumannii clinical isolates. Canadian Journal of Microbiology, 62(9) 794-801.
Puttilerpong, C., Chawanasit, W.,Laohawaleesan, W, Rungsang, W., Ritteeverkul, P. & (2011). Antimicroial use in hospital-acquired pneumonia with multidrug-resistant Acinetobacter baumannii at King Chulalongkorn Memorial Hospital. Thai Pharmaceutical and Health Science Journal, 6(1), 32-38. (in Thai)
Wieczorek, P., Sacha, P.,.Hauschild, T.,Zórawski, M., Krawczyk, M., & Tryniszewska, E. (2008). Multidrug resistant Acinetobacter baumannii-the role of AdeABC (RND family) efflux pump in resistance to antibiotics. Foloa Histichemica et Cytochemica, 46(3), 257-267.
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Published
2021-01-05
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