scholarly journals Plasmid-mediated Quinolone Resistance Genes in Salmonella typhi from Patients Attending Selected General Hospitals in Abuja Municipal, Nigeria

2020 ◽  
pp. 45-59
Author(s):  
R. Fasema ◽  
B. E. Bassey ◽  
Y. B. Ngwai ◽  
I. H. Nkene ◽  
R. H. Abimiku ◽  
...  
Keyword(s):  
General Hospital ◽  
Salmonella Typhi ◽  
Quinolone Resistance ◽  
General Hospitals ◽  
Microbiological Methods ◽  
Antimicrobial Resistance Profile ◽  
Mar Index ◽  
Stool Samples ◽  
Pcr Method ◽  
Polymerase Chain

This study investigated the antimicrobial resistance profile and presence of plasmid-mediated quinolone resistance (PMQR) genes in Salmonella typhi from patients attending selected general hospitals in Abuja municipal, Nigeria. Four hundred stool samples from patients with suspected typhoid fever were collected from Asokoro General Hospital Abuja (AGH), Garki Hospital Abuja (GHA), Maitama General Hospital Abuja (MGHA) and Wuse General Hospital Abuja (WGHA) and S. typhi was isolated and identified using standard microbiological methods. Antimicrobial susceptibility testing of the isolates was carried out using Clinical and Laboratory Standards Institute (CLSI) method. Molecular detection of PMQR genes in the ciprofloxacin-resistant isolates was carried out using Polymerase Chain Reaction (PCR) method. The overall occurrence of the isolates was 13.3% (53/400), with the highest hospital-related occurrence at WGHA (18.0%). The occurrence was highest at age 21-30yrs in AGHA (20.0%), GHA (33.3%) and WGHA (45.0%). The occurrence was higher in females at AGHA (12.7%) and GHA (16.0%); but higher in males at MGHA (11.4%) and WGHA (18.2%). Resistance to ciprofloxacin was the least at 30.2%, distributed as follows: AGHA (20.0%), GHA (35.7%), MGHA (36.4%) and WGHA (27.8%). The most common resistance phenotype was: NA-S-XT-AMC-TE-CRO-C-CN with overall occurrence of 9.4% (5/53) observed in AGH (10.0%), GHA (16.7%) and MGHA (18.2%) but not in WGHA. All (100%) isolates were multiple antibiotic resistance (MAR) isolates, with MAR indices above 0.2; and the commonest MAR index of 0.6 (30.0%) in AGHA, 0.8 (35.7%) in GHA; 0.8 (45.6%) in MGHA, and was 0.7 (38.9%) in WGHA. Multidrug resistance (MDR) was the commonest at 96.2% (51/53), with occurrences in the selected hospitals as follows: AGHA (90.0%), GHA (100.0%) and MGHA (100.0%) and WGHA (94.4%).The PMQR genes detected had overall frequency in the order: aac(6′)-Ib-cr (50.0%) >qnrB (37.5%) >qnrS (18.8%); qnrS was absent in AGHA and WGHA. The genes co-existed with one another with the qnrB+ aac(6′)-Ib-cr combination, present in isolates from all the hospitals, being the most common at (31.3%). Ciprofloxacin was the most effective antibiotic against the isolates; most isolates were MAR with prior exposure to antibiotics; most isolates were MDR and the ciprofloxacin-resistant isolatesharbored qnrS, qnrB and aac(6′)-Ib-cr PMQR genes, with aac(6′)-Ib-cr being the most prevalent.

Identification of Bacteroides forsythus in Subgingival Dental Plaque with the Aid of a Rapid PCR Method

1997 ◽  
Vol 76 (7) ◽  
pp. 1376-1380 ◽  
Author(s):  
J.H. Meurman ◽  
J. Wahlfors ◽  
A. Korhonen ◽  
P. Alakuijala ◽  
P. Väisänen ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Subgingival Plaque ◽  
Chain Reaction ◽  
Rapid Pcr ◽  
Microbiological Methods ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Pre Treatment ◽  
Culture Positive ◽  
Positive Results

Bacteroides forsythus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well-suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria.

scholarly journals Molecular Detection of Plasmid-Mediated Quinolone Resistance in Ciprofloxacin-Resistant Escherichia coli from Urine of Patients attending Garki Hospital, Abuja, Nigeria

2020 ◽  
Vol 1 (4) ◽  
Author(s):  
Moses Oghenaigah Eghieye ◽  
Istifanus Haruna Nkene ◽  
Rejoice Helma Abimiku ◽  
Yakubu Boyi Ngwai ◽  
Ibrahim Yahaya ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Molecular Detection ◽  
Quinolone Resistance ◽  
Chain Reaction ◽  
E Coli ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections

Urinary tract infections (UTIs) caused by Escherichia coli (E. coli) is common worldwide; and its successful treatment using antibiotics is limited by acquisition of resistance by the bacteria. This study investigated the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in ciprofloxacin-resistant E. coli from urine of patients with suspected cases of UTIs attending Garki Hospital Abuja (GHA), Nigeria. A total of 8 confirmed ciprofloxacin-resistant E. coli was screened for carriage of PMQR genes using polymerase chain reaction (PCR) method. The occurrences of the PMQR genes detected were in the order: aac-(6′)-Ib-cr (87.5%) > qnrB (50.0%) > qnrS (37.5%) > oqxAB (12.5%) > qnrA(0.0%). qnrB and qnrS did not exist alone, but in combination with other genes; aac-(6′)-Ib-crexisted both alone and in combination with others; the most prevalent patterns of existence were aac-(6′)-Ib-cr alone and aac-(6′)-Ib-cr + qnrB + qnrS at 25.0% each. This study has shown that the ciprofloxacin-resistant E. coli harbored aac-(6′)-Ib-cr, qnrB, qnrS and oqxAB PMQR genes, with aac-(6′)-Ib-cr being the most prevalent. The genes were present either alone or in combination with one another. This has implication for the clinical application of fluoroquinolones to treat UTI in the study location and environs. 

scholarly journals Comparative Study of Eleven Mechanical Pretreatment Protocols for Cryptosporidium parvum DNA Extraction from Stool Samples

2021 ◽  
Vol 9 (2) ◽  
pp. 297
Author(s):  
Laure Claudel ◽  
Nicolas Valeix ◽  
Louise Basmaciyan ◽  
Bruno Pereira ◽  
Damien Costa ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Cryptosporidium Parvum ◽  
Mechanical Pretreatment ◽  
Extraction Step ◽  
Physicochemical Features ◽  
Stool Samples ◽  
Pcr Method ◽  
Cryptosporidium Parvum Oocyst ◽  
Polymerase Chain ◽  
Commercial Kits

Nowadays, many commercial kits allow the polymerase chain reaction (PCR) detection of Cryptosporidium deoxyribonucleic acid (DNA) in stool samples, the efficiency of which relies on the extraction method used. Mechanical pretreatment of the stools using grinding beads has been reported to greatly improve this extraction step. However, optimization of this key step remains to be carried out. Indeed, many parameters could influence the pretreatment performances, among which the modulation of the speed and duration of the grinding step, in addition to the physicochemical features of the grinding beads, have never been evaluated to date. In this study, eleven commercial mechanical pretreatment matrixes (Lysis matrix tubes®, MP Biomedical, Irvine, CA, USA) composed of beads with different sizes, shapes, and molecular compositions, were evaluated for their performances in improving Cryptosporidium parvum oocyst DNA extraction before amplification by using our routinely used real-time PCR method. As expected, the eleven commercial mechanical pretreatment matrixes showed varying performances depending on the composition, size, and shape. All in all, the best performances were obtained when using the Lysing matrix, including ceramic beads with a median size (diameter of 1.4 mm).

scholarly journals Prevalence of Campylobacter spp. among imported broiler drumsticks and wings sold in Lithuania

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Dominyka Baltutytė ◽  
Laura Babonytė ◽  
Sigita Ramonaitė
Keyword(s):  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Chain Reaction ◽  
Microbiological Methods ◽  
Pcr Products ◽  
Pcr Method ◽  
Polymerase Chain ◽  
One Year ◽  
The One ◽  
Campylobacter Spp

The aim of this research was to estimate the prevalence of Campylobacter in imported broiler drumsticks and wings. During the one-year study period, 138 imported broiler samples (raw wings and drumsticks) were collected and tested from 3 different sellers. Campylobacter spp. were detected and isolated using traditional microbiological methods, identified using a multiplex polymerase chain reaction (PCR) method. The results of PCR products were analysed in agarose gel using electrophoresis. After an epidemiological study, C. jejuni and C. coli strains were selected and the prevalence of virulence genes was evaluated. The study identified Campylobacter spp. in 36 (26.1%) samples – 19 raw wings (27.9%) and 17 raw drumsticks (24.3%) samples were infected with these bacteria. Campylobacter spp. were most frequently detected in raw broiler samples during autumn (September–November) (47.2%) and winter (December–February) (41.6%) periods than spring (March–May) (5.5%) or summer (June–August) (5.5%). Contamination of products was not significantly impacted by the sale location (p > 0.05). The examination of virulence factors of Campylobacter spp. revealed that C. jejuni and C. coli strains contain 2 out of 3 virulence genes – CadF and CdtA. The CdtA gene was found in nearly all tested Campylobacter spp. strains isolated from broiler samples (94.4%).

Antimicrobial Resistance Profile and Quinolone Resistance Genes in Staphylococcus aureus from Patients Attending Federal Medical Centre Keffi, Nigeria

2020 ◽  
pp. 1-15
Author(s):  
E. A. Sunday ◽  
Y. B. Ngwai ◽  
R. H. Abimiku ◽  
I. H. Nkene ◽  
Y. Ibrahim ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Medical Centre ◽  
Clinical Samples ◽  
Resistance Profile ◽  
Antimicrobial Resistance Profile ◽  
Pcr Method

Aims: This study investigated the antimicrobial resistance profile and quinolone resistance genes in Staphylococcus aureus from patients attending Federal Medical Centre, Keffi, Nigeria. Methodology: A total of 240 clinical samples which comprised of high vaginal swabs, endocervical swabs, sputum, ear swabs, wound swabs, semen and eye swabs, were collected from the patients. Staphylococcus aureus was isolated and identified from these samples using standard microbiological method. Antimicrobial susceptibility testing of the isolates was performed and interpreted in accordance with the Clinical and Laboratory Standards Institute (CLSI) method. Ciprofloxacin-resistant S. aureus were screened for quinolone resistance genes using Polymerase Chain Reaction (PCR) method. Results: Out of 240 clinical samples, the prevalence of S. aureus was 21.3%. The prevalence in relation to clinical samples was higher in eye swab (45.5%) and ear swab (44.4%), but lower in sputum (14.5%). The isolates were more resistant to oxacillin (88.2%), sulphamethoxazole/ trimethoprim (82.4%) and erythromycin (76.5%), but less resistant to ciprofloxacin (19.6%) and levofloxacin (5.9%). The most common resistance phenotypes in the isolates were sulfamethoxazole/trimethoprim (SXT) - clindamycin (DA) – ofloxacin (OX) - erythromycin (E) - rifampicin (RD) and SXT-DA-OX-E- streptomycin (S) -RD with an occurrence of (13.7%) each. The percentage occurrences of multidrug resistant and extensive-drug resistant isolates were 92.2% and 7.8% respectively. The occurrences of quinolone resistance genes in the ciprofloxacin-resistant isolates were: aac(6′)-Ib-cr (60.0%), gyrA and gyrB (50.0%), parC (40.0%), qnrB (20.0%) and qnrS (10.0%). Conclusion: The isolates were less resistant to levofloxacin, cefoxitin, ciprofloxacin and gentamicin in the study location. Most of the ciprofloxacin-resistant isolates harbored quinolone resistance genes with aac(6′)-Ib-cr as the most common.

scholarly journals Detection of Flagellin Gene of Enteric Isolates of Salmonella typhi Using PCR in Baghdad Governorate

2019 ◽  
Vol 30 (3) ◽  
pp. 9
Author(s):  
Saba Saadoon Khazaal
Keyword(s):  
Polymerase Chain Reaction ◽  
Teaching Hospital ◽  
Virulence Gene ◽  
Salmonella Typhi ◽  
Chain Reaction ◽  
Flagellin Gene ◽  
Stool Samples ◽  
Polymerase Chain

The study was conducted to study the virulence gene (Flagellin gene) of Salmonella typhi. Stool samples were collected from adults patients, their age ranging from (15 – 60 ) years of the both sexes who suffering from diarrhea and visited AL- Yrmouk Teaching Hospital in Baghdad Governorate for the period between March to September 2017. The samples were transferred in cold boxes to the laboratory. In order to confirm the diagnosis, the polymerase chain reaction (PCR) was tested to diagnose the Flagellin gene responsible for the virulence of the bacteria. We conclude from the present study that flagellin gene in S. typhi is responsible for pathogenicity in this bacteria.

scholarly journals Antimicrobial Resistance Profile and Detection of Extended Spectrum and Amp C β-Lactamase Resistance Genes in Escherichia coli Isolated from Diarrheic Children in Lafia, Nasarawa State, Nigeria

2020 ◽  
pp. 15-27
Author(s):  
A. Zakou ◽  
I. H. Nkene ◽  
R. H. Abimiku ◽  
I. Yahaya ◽  
B. E. Bassey ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Medical Centre ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Extended Spectrum ◽  
Microbiological Methods ◽  
Antimicrobial Resistance Profile ◽  
Amoxicillin Clavulanic Acid ◽  
Pcr Method

Aims: This study evaluated the presence of extended spectrum β-lactamase (ESBL) and AmpC β-lactamase resistance genes in E. coli from stool of diarrheic children in some hospitals in Lafia metropolis, Nigeria. Methodology: A total of 70 stool samples of children were obtained from Dalhatu Araf Specialist Hospital, Lafia, M & D Hospital, Olivet Medical Centre and Sandaji Medical Centre, Lafia. Escherichia coli were isolated and identified using standard microbiological methods. Antimicrobial susceptibility of the isolates was tested using Clinical and Laboratory Standards Institute (CLSI) method. The phenotypic detection of ESBL and AmpC β-lactamase production in some antibiotic resistant isolates were carried out using disc method. The molecular detections of ESBL and AmpC resistance genes were carried out using Polymerase Chain reaction (PCR) method. Results: Of the 70 samples, the occurrence of E. coli was 100%. The isolates were highly resistant to ampicillin (97.14%), ciprofloxacin (90.00%), sulfamethoxazole/trimethoprim (84.29%), streptomycin (78.57%), amoxicillin/clavulanic acid (70.00%); moderate to gentamicin (38.57%), ceftazidime (37.14%) and cefotaxime (30.00%); and less resistant to cefoxitin (15.71%) and imipenem (8.57%). Twenty-one (30.00%) isolates were jointly resistant to both cefotaxime and ceftazidine. Of this number, 66.67% (14/21) were phenotypically confirmed ESBL producers; and the occurrences of ESBL resistance genes were: 7.14% (SHV), 42.86% (CTX-M) and 50.00% (TEM). Out of 11isolates resistant to cefoxitin, 4(36.36%) were phenotypically confirmed as AmpC β-lactamase producers; and the occurrence of AmpC genes were: 50.00% (CIT), 25.00% (FOX) and 25.00% (MOX). Conclusion: The isolates were least resistant to imipenem and cefoxitin and highly resistant to ampicillin, ciprofloxacin and sulfamethoxazole/trimethoprim. TEM and CTX-M ESBL genes were more frequent than SHV. CIT AmpC gene was more frequent than FOX and MOX.

scholarly journals Antimicrobial Resistance Profile of Escherichia coli Isolated from Urine of Patients in Selected General Hospitals in Abuja Municipal, Nigeria

2018 ◽  
pp. 1-10
Author(s):  
M. O. Eghieye ◽  
S. M. Jodi ◽  
B. E. Bassey ◽  
I. H. Nkene ◽  
R. H. Abimiku ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Clavulanic Acid ◽  
General Hospital ◽  
General Hospitals ◽  
Resistance Profile ◽  
E Coli ◽  
Antimicrobial Resistance Profile ◽  
Amoxicillin Clavulanic Acid

This study investigated the antimicrobial resistance profile of Escherichia coli from urine of patients with suspected urinary tract infections (UTIs) in selected general hospitals in Abuja Municipal, Nigeria. Four Hundred and Thirty urine samples were collected between September 2017 and May 2018 from patients attending Asokoro General Hospital (AGH), Garki Hospital Abuja (GHA) and Wuse General Hospital (WGH); and E. coli was isolated and identified by culture, microscopy and biochemical tests. The overall occurrence of E. coli was 52 (12.1%). The occurrences in relation to the hospitals were of the order: GHA (14.7%) > WGH (12.6%) > AGH (9.0%). The highest (50%) occurrence was at age 41-50 years in WGH, and the lowest (4.3%) was at age 31-40 years in AGH. More females than males harboured the bacteria in all the hospitals. Isolates from AGH showed highest (100.0%) resistance to Sulphamethoxazole/Trimethoprim but least (0.0%) resistance to Ciprofloxacin. Isolates from GHA showed the highest resistance to Cefotaxime and Streptomycin (95.2%) but least (23.8%) to Gentamicin and Imipenem. Isolates from WGH showed highest (88.8%) resistance to Amoxicillin/Clavulanic Acid but least (16.7%) to Sulphamethoxazole/Trimethoprim. The commonest antibiotic resistance phenotype in AGH was Amoxicillin/Clavulanic Acid-Streptomycin-Cefotaxime-Ceftazidime-Imipenem-Ampicillin (3.9%); in GHA was Amoxicillin/Clavulanic Acid-Streptomycin-Sulphamethoxazole/Trimethoprim-Cefotaxime-Ceftazidime-Ampicillin (7.7%); and in WGH was Amoxicillin/Clavulanic Acid-Streptomycin-Cefotaxime-Cefotaxime-Ceftazidime-Imipenem-Ampicillin (3.9%). All the isolates had MAR indices above 0.2; the most common index in AGH was 0.4 (at 30.8%), GHA was 0.7 (at 33.3%) and WGH was 0.7 (at 27.8%). The commonest class of antibiotic resistance was MDR with the order of occurrence as: GHA (92.2%) > WGH (77.7%) > AGH (76.6%). Ciprofloxacin, gentamicin and imipenem were the most effective antibiotics in the study location. However, MAR indices in this study have shown that the isolates originated from an environment where antibiotics are freely available and misused/abused. Hence, there is a need for greater monitoring of antibiotic supplies and use.

scholarly journals Comparison of Molecular and Parasitological Methods for Diagnosis of Human Trichostrongylosis

2021 ◽  
Vol 11 ◽  
Author(s):  
Mehdi Pandi ◽  
Meysam Sharifdini ◽  
Keyhan Ashrafi ◽  
Zahra Atrkar Roushan ◽  
Behnaz Rahmati ◽  
...  
Keyword(s):  
Agar Plate ◽  
Rural Populations ◽  
Diagnostic Specificity ◽  
Pcr Assay ◽  
Northern Iran ◽  
Wet Mount ◽  
Stool Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Fresh Stool

Human trichostrongyliasis is a zoonotic disease that is prevalent among rural populations in some countries. This study was performed to evaluate various parasitological methods and polymerase chain reaction (PCR) for the diagnosis of human trichostrongyliasis. A total of 206 fresh stool samples were collected from residents of endemic villages of Northern Iran. All samples were examined using conventional parasitological methods, including wet mount, formalin ethyl acetate concentration (FEAC), agar plate culture (APC), Harada–Mori culture (HMC), and Willis, along with the PCR technique. Among the total of 206 individuals examined, 72 people (35%) were found infected with Trichostrongylus species using combined parasitological methods. By considering the combined results of parasitological methods as the diagnostic gold standard, the Willis technique had a sensitivity of 91.7% compared with 52.8% for the APC, 40.3% for the HMC, 37.5% for FEAC, and 5.6% for the wet mount technique. The diagnostic specificity of all the parasitological methods was 100%. Furthermore, the PCR method detected Trichostrongylus spp. DNA in 79 fecal samples (38.3%) with a sensitivity of 97.2% and a specificity of 93.3%. According to the current findings, the Willis method was more sensitive than are the other parasitological methods in the diagnosis of human trichostrongyliasis. However, the PCR assay was more sensitive and more reliable in the detection of human trichostrongyliasis in comparison with the parasitological methods.

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