Molecular Characterization of Extended-Spectrum Beta-Lactamases in Escherichia coli and Klebsiella pneumoniae in Accra, Ghana

Extended-spectrum beta-lactamases (ESBLs) are plasmid-mediated beta lactamases that are capable of hydrolysing beta-lactams except carbapenems and cephamycins. The ESBL types include SHV, TEM and CTX-M, OXA, PER and VEB-1. The most common ones isolated from clinical specimen are the CTX-M, SHV and TEM.  The specific ESBL-producing organisms have different genetic characteristics which mark their identification at the molecular level. This work sought to determine the genetic characterization of ESBL-producing K. pneumoniae and E. coli in Accra. The molecular investigations of the ESBL-coding genes included extraction of 100 DNA templates of phenotypic ESBL-producing isolates by boiling method, preparation of the PCR reaction mixture using appropriate primers, standard PCR reaction in a thermocycler,   agarose gel electrophoresis, bands visualization by ultraviolet trans-illumination and bands photography using a Kodak EDAS 290 gel documentation system. The results significantly (p<0.05) indicated that of the 100 ESBL producers, 90(90%) possess CTX-M genes and 25(25%) had TEM genes. None of the ESBL producers possesses SHV genes. Seventy (70%) of the ESBL producers possess only CTX-M genes and 5(5%) had only TEM genes. Twenty (20%) of the isolates had both CTX-M and TEM genes. Of the 100 ESBL phenotypes, 78(78%) and 2(2%) were positive for CTX-M-1group and CTX-M-9group ESBL genes respectively. Organisms producing CTX-M-type ESBL are more prevalent in Accra than other ESBL types. CTX-M-1group producing isolates dominated the ESBL phenotypes with CTX-M-15 likely to be the dominate CTX-M-type ESBL. There is the need for further studies into the characteristic transmission, pathogenesis, antibiotic resistance expression, and infection control of CTX-M-type ESBL and TEM-type ESBL in Accra. Keywords: Extended spectrum beta-lactamase, CTX-M genes, TEM genes, SHV genes, Molecula

Detection of Human Papillomavirus Genotypes and Epstein-Barr Virus in Nasopharyngeal Carcinomas at the Korle-Bu Teaching Hospital, Ghana

Nasopharyngeal carcinomas (NPC) are endemic in Far East Asia and commonly harbour Epstein-Barr virus (EBV) which is known to serve as a key oncogenic promoter. Human papillomavirus (HPV) is known to contribute to the pathogenesis of NPC. However, in Ghana these two viruses have not been linked to NPC prevalence. This study was designed to determine the HPV genotypes and EBV involved in NPC tissue biopsies. A retrospective study design involving 72 formalin-fixed paraffin-embedded tissue (FFPET) samples of NPC from 2006 to 2012 were retrieved from the Department of Pathology, University of Ghana School of Biomedical and Allied Health Sciences. Sections were taken for histological analysis and for DNA lysate preparation. The DNA lysates were subjected to polymerase chain reaction (PCR) analysis to determine the presence of HPV genotypes and EBV. HPV specific primers were used to type for fourteen HPV genotypes (HPV-16, 18, 6/11, 31, 33, 35, 44, 42, 43, 45, 56, 52, 58, and 59). Out of the 72 NPC biopsies analyzed by PCR, EBV DNA was present in 18 (25%) cases and HPV DNA in 14 (19.23%). High risk HPV (HR-HPV) genotypes 18 and 31 were associated with the NPC. There were 3 (4.2%) cases of coinfection by both viruses. The EBV DNA present in the undifferentiated variant of the NPC and the histopathology of the NPC in Ghana is similar to the type described in endemic areas

HER-2 Protein Overexpression in Patients with Gastric and Oesophageal Adenocarcinoma at a Tertiary Care Facility in Ghana

The prognosis of gastric and oesophageal adenocarcinoma remains generally poor. However, mounting evidence suggests a positive role of human epidermal growth factor receptor-2 (HER-2) expression in the prognosis of patients with these cancers. In this work, the patterns of HER-2 protein expression were determined in patients with gastric or oesophageal adenocarcinoma. Retrospectively, we reviewed records of gastric and oesophageal biopsies received from 2008 to 2012 and their corresponding archived formalin-fixed paraffin-embedded tissue blocks selected for immunohistochemical analysis. The prevalence of gastric and oesophageal adenocarcinomas and their association with HER-2 protein overexpression were evaluated. Gastric adenocarcinoma made up 18.79% of the gastric biopsies reviewed, and majority of these cancers occurred in males. Regarding the tumour type, HER-2 overexpression was common in the intestinal subtype compared to the diffuse type. Although squamous cell carcinoma was observed to be the commonest (31%) tumour type in the oesophagus compared to adenocarcinoma (8.79%), HER-2 was overexpressed in 42.9% of oesophageal adenocarcinomas, like gastric adenocarcinoma (41.4%). There is a high prevalence of gastric and oesophageal adenocarcinoma, with significant overexpression of HER-2 in these tumours, a window of hope for the management of patients with these cancers

Malaria elimination in Ghana:recommendations for reactive case detection strategy implementation in a low endemic area of Asutsuare, Ghana

Background: Progress toward malaria elimination is increasing as many countries near zero indigenous malaria cases. In settings nearing elimination, interventions will be most effective at interrupting transmission when targeted at the residual foci of transmission. These foci may be missed due to asymptomatic infections. To solve this problem, the World Health Organization recommends reactive case detection (RACD). This case study was conducted to identify individuals with asymptomatic malaria, their predisposing risk factors and recommend RACD in Asutsuare, Ghana based on literature review and a cross sectional study. Methods: The study involved a search on PubMed and Google Scholar of literature published between 1st January, 2009–14th August, 2023 using the search terms “malaria” in “Asutsuare”. Furthermore, structured questionnaires were administered to one hundred individuals without symptoms of malaria and screened using rapid diagnostic test (RDT) kits, microscopy and real-time polymerase chain reaction (rt-PCR). Malaria prevalence based on the three diagnostic techniques as well as potential malaria risk factors were assessed through questionnaires in a cross-sectional study. Results: Cumulatively, sixty-four (64) studies (Google Scholar, 57 and PubMed, 7) were reviewed and 22 studies included in the literature on malaria in Asutsuare, Ghana. Significant risk factors were occupation, distance from a house to a waterbody, age group and educational level. Out of the 100 samples, 3 (3%) were positive by RDT, 6 (6%) by microscopy and 9 (9%) by rt-PCR. Ages 5–14.9 years had the highest mean malaria parasite densities of 560 parasites/µl with Plasmodium falciparum as the dominant species in 4 participants. Moreover, in the age group ≥ 15, 2 participants (1 each) harboured P. falciparum and Plasmodium malariae parasites. RDT had a higher sensitivity (76.54%; CI95 66.82–85.54) than rt-PCR (33.33%; CI95 4.33–77.72), while both rt-PCR and RDT were observed to have a higher specificity (92.55; CI95 85.26–96.95) and (97.30; CI95 93.87–99.13), respectively in the diagnosis of malaria. Conclusion: In Asutsuare, Ghana, a low endemic area, the elimination of malaria may require finding individuals with asymptomatic infections. Given the low prevalence of asymptomatic individuals identified in this study and as repleted in the literature review, which favours RACD, Asutsuare is a possible setting receptive for RACD implementation.</p

Characterization of Campylobacter associated gastric enteritis among patients with Human Immunodeficiency Virus (HIV) in a hospital in Accra, Ghana.

BackgroundCampylobacter infections in HIV positive patients often present with substantial mortality and morbidity when compared to HIV negative patients.AimThis study assessed the prevalence of Campylobacter, antibiotic resistance phenotypes and genetic factors, and risk of Campylobacter infection associated with living in close proximity to domestic animals in HIV patients with gastric enteritis at Korle-Bu Teaching Hospital, Accra, Ghana.MethodsResistance to different antibiotics was assessed with Kirby-Bauer disk diffusion method. In addition, all the Campylobacter isolates were tested for ampicillin (blaOXA-61), erythromycin (aph-3-1), tetracycline tet(O), streptomycin (aadE), and the energy-dependent multi-drug efflux pump (cmeB) resistance genes using multiplex polymerase chain reaction.ResultsOut of a total of 140 (97 females and 43 males) tested patients, 71 (50.7%) patients were positive for Campylobacter coli. Female patients aged within 31-40 years (31.6%) and 41-50 years (31.6%) had high frequency of Campylobacter infection. Most of the infected patients lived in close proximity to chickens (53.5%), however, some patients (14.1%) lived in close proximity to goats. Phenotypic resistance evaluation revealed widespread resistance to ampicillin (100%), tetracycline (100%), ciprofloxacin (71.8%), erythromycin (69%), and gentamicin (49.3%). However, limited no of isolates contained blaOXA-61 (1.41%), cmeB (7.0%) and tet(O (7.0%) resistance genes.ConclusionHIV patients with gastric enteritis were infected with resistant Campylobacter coli. Further studies are required to examine correlation of infected patients with C. coli and risk of living in close proximity to poultry birds. There is the need for routine investigation of Campylobacter in patients with gastroenteritis in order to assist in the development of strategies for combating diseases involving resistant zoonotic bacteria strains

Antiplasmodial and Genotoxic Study of Selected Ghanaian Medicinal Plants

Ethnopharmacological Relevance. Development of resistance to antimalarial drugs by Plasmodium falciparum is still rampant, and there is an urgent need for novel drugs to either standalone or to partner artemisinin for treatment of malaria. Traditionally, plants have, over the years, been a good source of antimalarial drugs. Efficacy and safety of such plants need to be scientifically authenticated. Aims, Materials, and Method. This study investigated the in vitro antiplasmodial activity, cytotoxicity, and genotoxicity of aqueous extracts of Acanthospermum hispidum DC, Alstonia boone (De Wild), Cocos nucifera L, Cymbopogon citratus (DC.) Stapf, Morinda lucida Benth, Psidium guajava, Phyllanthus niruri L, and Senna siamea Lam. Results. Five out of the eight plants, A. boonei stem bark, S; siamea Lam root, M. lucida Benth leaves, P. niruri, and A. hispidum DC whole plants, showed varying degrees of antiplasmodial activity against the asexual stage of the parasite. The most active extract against chloroquine-sensitive (3D7) and chloroquine-resistant (Dd2) P. falciparum strains is the A. hispidum extract which yielded a mean inhibitory concentration at 50% (IC50) of 3.66 µg/ml and 3.71 µg/ml for 3D7 and Dd2, respectively. This was followed by S. siamea Lam with 3.95 µg/ml for 3D7 and 4.47 µg/ml for Dd2. The IC50 values of the A. boonei extract against 3D7 and Dd2 P. falciparum parasites were 5.13 µg/ml and 3.62 µg/ml, respectively. For the M. lucida Benth extract, the least IC50 value was 6.46 µg/ml. All five extracts exhibited dose-dependent antiplasmodial activity. Assessment of the genotoxic effects the A. hispidum extract by the comet assay revealed substantial damage to P. falciparum DNA. Conclusion. This study demonstrates that the crude extract of A. hispidum DC, one of the plants used traditionally to treat malaria, inhibits the growth of P. falciparum in vitro and could be a potential source of antimalarial drug. The report has highlighted genotoxic and cytotoxic effects of the selected plant extracts on human leukocytes as well

SERO-prevalence of herpes simplex virus type 1 and type 2 among women attending routine Cervicare clinics in Ghana

Abstract Background Herpes simplex virus infection is a global health concern with disproportionately high burden in low and middle-income countries. There is a paucity of data on the prevalence of HSV infection in Ghana, which necessitated the present study. The aim of the study was to provide up-to-date data on sero-prevalence of HSV-1 and HSV-2 infection among women attending Cervicare clinics in Ghana. Methods This was a cross-sectional study in which 380 women attending routine Cervicare clinics at Regional Hospitals in Kumasi and Accra, Ghana were enrolled into the study. Serum HSV-1 IgG and HSV-2 IgG were determined by ELISA method. The Chi-square test was used to investigate the association between sero-prevalence of HSV-1 and HSV-2 and socio-demographic and behavioral factors using the Statistical Package for the Social Scientists (SPSS) version 22. Statistical significance was accepted at p < 0.05. Results The overall HSV-1 and HSV-2 sero-prevalence estimates were 99.2% (95% CI: 98.0–100%) and 78.4% (95% CI: 74.5–81.8%) respectively. The study observed 78.2% cross-positive prevalence of HSV-1 and HSV-2 among the studied participants. There was no association between the presence of HSV-1 and HSV-2 infection and age (χ2 = 2.351, p = 0.799 and χ2 = 1.655, p = 0.895 respectively). Our findings however, revealed association between the prevalence of HSV-2 and the age at coitarche (p = 0.021) as well as with number of sexual partners (p = 0.022). Conclusions The sero-prevalence estimates of HSV-1 and HSV-2 among the study population of women in Ghana were found to be high. This high prevalence could be attributed to high endemicity and inadequate intervention in this population. There is the need to raise awareness through organized public health screening and education to ensure control

Trichomonas vaginalis infection and the diagnostic significance of detection tests among Ghanaian outpatients

Abstract Background There is little data on Trichomonas vaginalis infection in Ghana. This study evaluated the prevalence of trichomoniasis using different diagnostic methods and determined the risk factors for infection in patients. Methods A structured questionnaire was administered. Vaginal swabs, urethral swabs and urine specimens were obtained from consenting patients; and the samples processed following standard protocols. The presence of T. vaginalis was determined using wet mount microscopy and polymerase chain reaction (PCR) as gold standard. We also assessed the diagnostic performance the JD’s Trichomonas V® rapid antigen test to inform clinical practice. Results The PCR assay detected T. vaginalis positivity in 64 of 150 patients (42.6, 95%CI:35.0, 50.6) including all positive samples of wet mount microscopy and JD’s Trichomonas V® test. Wet mount microscopy showed low sensitivity (31.6%), high specificity (100%), moderate positive predictive value (75.0%), moderate positive likelihood ratio (3.0), and weak agreement (Cohen’s kappa, 0.283) with PCR assay. The JD’s Trichomonas V® test displayed lower sensitivity (25.0%), specificity (83.3%), and weaker measure of agreement (Cohen’s kappa, 0.233) with PCR. In multivariate analysis, the strongest independent predictor for T. vaginalis was female gender [adjusted odds ratio (AOR), 24.89; 95% confidence interval (CI): 10.58, 51.21; P-value< 0.001]. Knowledge of STI showed a protective effect against infection with the parasite (AOR, 0.13; 95%CI: 0.07, 0.29; P-value< 0.017). Conclusion The sensitivity of wet mount microscopy was low for T. vaginalis screening in our region. The JD’s Trichomonas V® test should not be considered as an alternative test. We recommend mandatory PCR assay for confirmation of negative wet mount results

Changing Patterns of Rotavirus Genotypes in Ghana: Emergence of Human Rotavirus G9 as a Major Cause of Diarrhea in Children

Genotyping of human rotaviruses was performed on 312 rotavirus-positive samples collected from 2,205 young children with diarrhea in the Upper East District of Ghana, a rural community. Of the 271 (86.9%) rotavirus strains that could be VP7 (G) or VP4 (P) characterized, 73 (26.9%) were of G9 specificity. The predominant G9 genotype was G9P[8], which constituted 79.5% of all G9 strains detected, followed by G9P[6] (12.3%), G9P[10] (2.7%), and G9P[4] (1.3%). G9 strains with mixed P types constituted 2.7% of all G9 strains found in the study. All the G9P[8] strains had a long RNA electrophoretic pattern with VP6 subgroup II specificity. Four G9 isolates, GH1319, GH1416, GH3550, and GH3574, which were selected based on the abundance of stool material and were representative of the three electropherotypes observed, were cloned and sequenced. The Ghanaian isolates shared more than 98% sequence nucleotide homology with other G9 strains from the United States (US1205), Malawi (MW69), Brazil (R160), Japan (95H115), and Nigeria (Bulumkutu). However, they showed only 95% nucleotide homology with the Thai G9 strain Mc345. Phylogenetic analysis of the nucleic acid sequence revealed the existence of at least three clusters, with Ghanaian strains forming one cluster, Nigerian and Brazilian strains forming a second cluster, and U.S., Malawian, and Japanese strains forming a third