PCR Amplification on a Microarray of Gel-Immobilized Oligonucleotides: Detection of Bacterial Toxin- and Drug-Resistant Genes and Their Mutations

Background: Acinetobacter baumannii is a gram-negative bacterium classified as an opportunistic pathogen in humans by the World Health Organization. Different genetic determinants contribute to multidrug resistance, and transform it as a nosocomial pathogen. Aim: Using in-silico PCR, this analysis aims to characterize the 13 distinct drug resistant genes found in 19 virulent A.baumannii strains. Materials & Methods: There were 11 A.baumannii multidrug resistance genes chosen. In-silico PCR amplification was performed using forward and reverse primers from the 11 genes described in previous research. The amplicon bands were detected in 19 strains of A.baumannii that were set as default on the server. Results: Among the 13 multidrug resistance genes studied, tet A, tet B, Sul 1, Sul 2, DfrA1, ISAba-1 and ISAba-125 were detected among the 19 virulent strains of A.baumannii. Conclusion: The findings of the study documents the frequency of tet A, tet B, Sul 1, Sul 2, DfrA1, ISAba-1 and ISAba-125 like from the selected strains of A. baumannii. However, more experimental validation is needed in order to conduct routine surveillance on drug-resistant A. baumannii strains in hospital settings.

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409 EFFICIENCY OF CO-TRANSFECTION OF TRANSGENE AND Neor GENE INTO GOAT AND PIG FETAL FIBROBLASTS

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab409 ◽

2007 ◽

Vol 19 (1) ◽

pp. 320

Author(s):

Y. M. Shin ◽

S. M. Chang ◽

B. C. Kim ◽

C. S. Park ◽

D. I. Jin

Keyword(s):

Transfection Efficiency ◽

Somatic Cells ◽

Pcr Amplification ◽

Pcr Analysis ◽

Fibroblast Cells ◽

Drug Resistant ◽

Fetal Fibroblast ◽

Resistant Genes ◽

G418 Selection ◽

Fetal Fibroblasts

Transgenic animals can be generated by nuclear transfer with genetically modified somatic cells in which the essential procedure of transgene transfection is required. Most transgene vectors are constructed to contain transgene and drug-resistant genes to enrich for somatic cells in which transgene integration has occurred. However, construction of transgene vectors along with drug-resistant genes may not be easy, due to inappropriate restriction sites. Therefore, in this study, two separate constructs, human tPA cDNA fused to β-casein promoter sequence as a transgene vector and neomycin-resistant gene (Neor) driven by PGK promoter as a drug-selectable gene, were co-transfected into pig and goat fetal fibroblast cells to estimate the efficiency of transgene transfection following G418 selection. First, goat fetal fibroblasts (GFF) and pig fetal fibroblasts (PFF) were tested for G418 resistance with different concentrations of G418. The pertinent concentrations of G418 were 800 µg mL−1 for GFF and 200 µg mL−1 for PFF. The linearized tPA vector and Neor gene vector were co-transfected into goat fetal fibroblasts and pig fetal fibroblasts with FuGENE6 transfection reagent (Roche Diagnostics, Mannheim, Germany). The cells were selected following exposure of 800 µg mL−1 and 200 µg mL−1 G418 for GFF and PFF, respectively, for 14 days. Cell colonies surviving G418 selection were assayed by PCR amplification with tPA-specific primers. Initially 2 × 106 GFF and PFF were transfected. Resistant colonies were counted and transferred to 24-well plates for expansion and PCR analysis. The results of co-transfection experiments are summarized in Table 1. The transfection of 2 × 106 GFF and PFF yielded an estimated 96 and 93 colonies, respectively, which survived as the G418 selection. However, 54 colonies of GFF and 39 colonies of PFF proliferated during expansion and were subjected to PCR analysis. Twenty-three and 5 of these colonies were identified to contain tPA transgene in GFF and PFF colonies, respectively. Transfection frequencies for tPA gene were 42.6% and 12.8% in GFF and PFF, respectively. These results suggest that co-transfection of transgene vector with Neor gene can be an alternative method for transfection of transgenes into fetal fibroblast cells. Table 1. Transfection efficiency of goat fetal fibroblasts (GFF) and pig fetal fibroblasts (PFF) following co-transfection of tPA gene and Neor gene

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Complete Genome Sequences of Four Extensively Drug-Resistant Acinetobacter baumannii Isolates from Thailand

Microbiology Resource Announcements ◽

10.1128/mra.00949-20 ◽

2020 ◽

Vol 9 (40) ◽

Author(s):

Peechanika Chopjitt ◽

Thidathip Wongsurawat ◽

Piroon Jenjaroenpun ◽

Parichart Boueroy ◽

Rujirat Hatrongjit ◽

...

Keyword(s):

Acinetobacter Baumannii ◽

Complete Genome ◽

Sequence Type ◽

Clinical Isolates ◽

Drug Resistant ◽

Genome Sequences ◽

Content Type ◽

Type Isolate ◽

Resistant Genes ◽

Extensively Drug Resistant

ABSTRACT Here, we report the complete genome sequences of four clinical isolates of extensively drug-resistant Acinetobacter baumannii (XDRAB), isolated in Thailand. These results revealed multiple antimicrobial-resistant genes, each involving two sequence type 16 (ST16) isolates, ST2, and a novel sequence type isolate, ST1479.

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Screening and cloning of multi-drug resistant genes in HL-60/MDR cells

Leukemia Research ◽

10.1016/j.leukres.2008.11.018 ◽

2009 ◽

Vol 33 (8) ◽

pp. 1120-1123 ◽

Cited By ~ 7

Author(s):

Gai-Huan Zheng ◽

Jin-Rong Fu ◽

You-Hua Xu ◽

Xian-Qing Jin ◽

Wen-Li Liu ◽

...

Keyword(s):

Drug Resistant ◽

Resistant Genes

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Detection of extensively drug-resistant and hypervirulent Klebsiella pneumoniae ST15, ST147, ST377 and ST442 in Iran

Acta Microbiologica et Immunologica Hungarica ◽

10.1556/030.2021.01562 ◽

2021 ◽

Author(s):

Sara Davoudabadi ◽

Hossein Goudarzi ◽

Mehdi Goudarzi ◽

Abdollah Ardebili ◽

Ebrahim Faghihloo ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Pcr Amplification ◽

Multidrug Resistant ◽

Diffusion Method ◽

Drug Resistant ◽

Carbapenem Resistant ◽

Extensively Drug Resistant ◽

String Test ◽

Pcr Method ◽

Extensively Drug Resistance

Abstract In this study, we focused on the emergence of extensively drug-resistant (XDR), pandrug-resistant (PDR), and hypervirulent Klebsiella pneumoniae (hvKP) in Iran. During 2018 to 2020 a total of 52 K. pneumoniae isolates were collected from different clinical specimens. The hvKP isolates were identified by PCR amplification of virulence and capsular serotype-specific genes. Hypermucoviscous K. pneumoniae (hmKP) were identified by string test. Carbapenem-resistant hvKP (CR-hvKP), multidrug-resistant hvKP (MDR-hvKP), extensively drug-resistant hvKP (XDR-hvKP), and pandrug-resistant hvKP (PDR-hvKP) were determined by disc diffusion method, Carba-NP test and PCR method. XDR-hvKP isolates were typed by multilocus sequence typing (MLST). Among all K. pneumoniae isolates 14 (26.9%) were identified as hvKP and 78.6% (11/14) of them were hmKP however, none of the classic K. pneumoniae (cKP) isolates were hmKP. The predominant capsular serotype of hvKP was K2 (42.85%) followed by K1 (35.71%). The prevalence of MDR-hvKP, XDR-hvKP and PDR-hvKP isolates were 6 (42.9%), 5 (35.7%) and 1 (7.1%), respectively. ESBL production was found in 85.7% of hvKP isolates and most of them carried bla TEM gene (78.6%) and 6 isolates (42.9%) were CR-hvKP. Among hvKP isolates, 1 (7.1%), 2 (14.3%), 3 (21.4%), 8 (28.6%), and 11 (78.6%) carried bla NDM-6, bla OXA-48, bla CTX-M, bla SHV, and bla TEM genes, respectively. According to MLST analysis, 2, 1, 1, and 1 XDR-hvKP isolates belonged to ST15, ST377, ST442, and ST147, respectively. The occurrence of such isolates is deeply concerning due to the combination of hypervirulence and extensively drug-resistance or pandrug-resistance.

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Graphdiyne-Based One-Step DNA Fluorescent Sensing Platform for the Detection of Mycobacterium tuberculosis and Its Drug-Resistant Genes

ACS Applied Materials & Interfaces ◽

10.1021/acsami.9b15248 ◽

2019 ◽

Vol 11 (39) ◽

pp. 35622-35629 ◽

Cited By ~ 8

Author(s):

Fan Chang ◽

Lijun Huang ◽

Chaozhong Guo ◽

Guoming Xie ◽

Jiaqiang Li ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Resistant ◽

Fluorescent Sensing ◽

Resistant Genes ◽

Sensing Platform ◽

One Step

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Comparative assessment on the prevalence of mutations in the Plasmodium falciparum drug-resistant genes in two different ecotypes of Odisha state, India

Infection Genetics and Evolution ◽

10.1016/j.meegid.2016.03.014 ◽

2016 ◽

Vol 41 ◽

pp. 47-55 ◽

Cited By ~ 6

Author(s):

Narayani Prasad Kar ◽

Kshipra Chauhan ◽

Nutan Nanda ◽

Ashwani Kumar ◽

Jane M. Carlton ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Comparative Assessment ◽

Drug Resistant ◽

Resistant Genes

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New Variant of mcr-3 in an Extensively Drug-Resistant Escherichia coli Clinical Isolate Carrying mcr-1 and blaNDM-5

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01757-17 ◽

2017 ◽

Vol 61 (12) ◽

Cited By ~ 37

Author(s):

Lu Liu ◽

Yu Feng ◽

Xiaoxia Zhang ◽

Alan McNally ◽

Zhiyong Zong

Keyword(s):

Escherichia Coli ◽

Clinical Isolate ◽

Amino Acid Substitutions ◽

Drug Resistant ◽

Content Type ◽

Carbapenem Resistant ◽

Resistant Genes ◽

Extensively Drug Resistant ◽

New Variant ◽

Carbapenemase Gene

ABSTRACT A colistin- and carbapenem-resistant Escherichia coli clinical isolate was found to carry two plasmid-borne colistin-resistant genes, mcr-1 and the newly identified mcr-3, and a carbapenemase gene, bla NDM-5. mcr-3 is a new variant (mcr-3.5) in the isolate and encodes three amino acid substitutions compared with the original MCR-3. mcr-3 was carried by a TnAs3-like transposon on a self-transmissible IncP plasmid in the isolate, highlighting that mcr-3 may have widely spread.

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