النشر العلمي

  • Detection of Plasmodium Falciparum Using Nested PCR Among Suspected Patients in Wad Medani City, Gezira State, Sudan

Introduction:
Plasmodium falciparum is a virulent species and causes most severe and pathogenic form of malaria, characterized by high parasitic burdens and vital organ dysfunctions. The diagnosis of falciparum malaria in Sudan depends on microscopy and Rapid diagnostic test (RDTs). Nested PCR targeted 18S rRNA gene provides an alternative technique for identification of parasite in thick and blood film.
Objective:
The study aims to estimate the diagnostic performance of 18S rRNA gene analysis by nPCR for detection of P. falciparum versus microscopic detection.
Method:
This is a cross sectional hospital based study. Two hundred and twenty blood samples and relevant data were collected from patients with falciparum malaria attending Wad Medani Teaching Hospitals and 26 samples from healthy participants. Parasite count was counted using stained thick blood film. The DNA extraction was done using TE buffer. Moreover, Nested PCR for 18S rRNA gene was done using specific primers. Data were analyzed using MedCalc programs (V. 16).
Results:
The microscopic examination of blood films showed that all patients (220/100%) were positive for P. falciparum among patients. one sample (3.8%) was positive among healthy participants (1/26). When using nPCR for 18S rRNA gene 219 samples were positive (99.5%) form patients and one sample was positive (3.8%) among healthy participants. The area under curve (AUC), sensitivity, specificity, positive predictive value, negative predictive sensitivity and Weighted Kappa agreement for the nPCR were 0.935, 98.6%, 88.6%, 60.1%, 99.7% and 0.871(95% CI:0.770 - 0.972), respectively.
Conclusion:
Nested PCR for 18S rRNA gene is sensitive, specific, reliable technique for diagnosis of Plasmodium falciparum in the endemic study area and can be introduced as a confirmatory diagnostic tool.

published in International Journal of Medical Science and Health Research

  • Evaluation of Loop Mediated Isothermal Amplification (LAMP) for Diagnosis of Plasmodium falciparum in Wad Medani City, Gezira State, Sudan

Plasmodium falciparum considered as the most serious form of species causes malaria compared with other species. Diagnosis of falciparum malaria in Sudan remain a major problem, thelaboratory diagnosis depends solely on microscopy and RDTs. Loop mediated isothermal amplification (LAMP) assay is a molecular technique done in isothermal temperature using simple, inexpensive instruments for detection of falciparum malaria. The aim of the study is to evaluate the
diagnostic performance of loop mediated isothermal amplification (LAMP) assay for detection of 
falciparum and compare with microscopic detection. A cross sectional hospital based study
conducted on 220 blood samples collected from participants suspected to have falciparum malaria
attending Wad Medani Teaching Hospitals and 26 healthy participants during the period November
2018 to January 2019. Thick blood films were done and used for P. falciparum detection. The
extracted DNA by TE buffer was amplified by LAMP assay targeting 18S rRNA gene. Data were
analyzed using Medical calculator (MedCalc) programs (V. 16). The results showed that the
sensitivity, specificity, positive predictive value, negative predictive values were 99.1%, 84.6%,
53.2%, 99.8% respectively. Validation of LAMP diagnostic performance revealed that area under
the curve is 0.919, while Weighted Kappa is 0.866. The study concluded that the LAMP assay had
the identical diagnostic performance compared with microscopy in diagnosis of Plasmodium
falciparum malaria. This gives a relative effortlessness application of LAMP assay in Sudan after
availing the required logistics.

published in International Journal of TROPICAL DISEASE & Health

  • Molecular Detection of H. Pylori from Gastric Biopsies of Dyspeptic Patients Attending Endoscopy Center, Gezira State, Sudan

Background: H. pylori are Gram negative bacteria cause most common of gastrointestinal tract infections worldwide. H. pylori can lead to serious symptomic or a symptomic illness including ulcers, gastritis, doudenitis, oesophagitis. Serious complications like gastric atrophic, mucosa-associated lymphoid tissue (MALT) lymphoma can occur. H. pylori posses cag A gene which is a virulent factor and marker for the pathogenic strain. This strain associated with greater inflammations and increased the risk of developing both peptic ulcer diseases and gastric carcinoma.
Objective: The study conducted to focus on detection of H. pylori and cag A gene using polymerase chain reaction (PCR) method in biopsy samples from upper gastrointestinal diseases patients in Gezira State, Sudan.
Materials and Methods: Descriptive cross - sectional study was carried out during 2016 – 2019 in Gezira State, Sudan. A total of 102 antrum biopsy samples were collected from adult male and female, their age between (20 – 70 years). Biopsy sample collected by gastroenterologists at Gezira Center for G.I.T. Endoscopy and Laparoscopic Surgery. H. pylori DNA extracted to apply the PCR technique in order to investigate H. pylori infection and cag A gene. In this study PCR for 16s rRNA accepted as gold standard method to identify the H .pylori.
Results: In 102 adult dyspeptic patients (45% male, 55% female, mean of age 46.1 ± 13 years). H. pylori was detected in 53 (51.9%) biopsy samples using PCR 16s rRNA, 22 (41.5%) were positive for cag A. Epigastric pain was a common clinical feature in individuals infected with H. pylori 71 (74%), dyspepsia 28 (29.5%) and vomiting 30 (25.8%). H. pylori infection predominant in gastritis and ulcer patients. Frequency of H. pylori positive cag A was common among in ulcers, gastritis, duodenitis and esophagitis patients. Out of 102 patients 9% reported mass endoscopy finding from them 56% infected with H. pylori when tested by the PCR in those positive H. pylori 80% reported positive cag A.
Conclusion: Frequency of H. pylori infection using 16s rRNA is 53 (51.9%) predominant in ulcers and gastritis patients. Cag A gene highest in gastritis and ulcers patients compare with other diseases, this gene play a role for determination the clinical outcome of H. pylori infection.

published in EUROPEAN ACADEMIC RESEARCH

  • The Role of Anti Mullerian Hormone (AMH) Levels as Predictive and Diagnostic Biomarker of Age at Menopause Among Infertile Sudanese Women

Abstract: Infertility is a global health problem with an increasing incidence rate among Sudanese women. The study aimed to compare
serum anti-mullerian hormone (AMH), folic stimulating hormone (FSH) and luteinizing hormone (LH) between primary infertility and
secondary infertility. Furthermore, also to study the effect of woman's age on these hormones. The study was carried out in Khartoum
State, Sudan during the period from January to March 2017. A cross sectional hospital included 260 infertile women [169 with primary
infertility (mean age 32.7 ± 7.26 years) and 91 with secondary infertility (mean age 35.02 ± 6.25 years). Three ml of venous blood were
collected in plain container and serum were obtained by centrifugation, all serum samples were analyzed for AMH, FSH and LH
concentration using ELISA techniques. The results were showed significant difference in LH and AMH between primary infertility and
secondary infertility (P value = 0.017, 0.001 respectively) and insignificant differences in FSH between them (P value = 0.606).
Furthermore, the serum AMH level decreased in elder ages (P value = 0.000; r = - 0.4564) and increased of FSH level (P value =
0.001; r = - 0.199). In addition, the serum FSH levels increase in elder ages (P value = 0.001: r = -0.2036). In conclusion, the results of
this study showed that hormones play a crucial role in diagnosis infertile women and serum AMH concentration has created more
promising results to predicting age at menopause.

published in International Journal of Academic Health and Medical Research (IJAHMR)

  • The Role of TNF-α Levels as Predictive Diagnostic Biomarker Among Children with Severe Falciparum Malaria in Endemic Area in Sudan

Abstract: Tumor necrosis factor alpha (TNF-α) level is a central proinflammatory cytokine. Their production associated with severe
falciparum malaria. The purpose of this study was to compare TNF-α levels between severe falciparum malaria, uncomplicated malaria
and control. Furthermore to evaluate the association between TNF-α levels and malaria parasitemia and malaria parasite count. A case
control hospital based study was included 300 Sudanese children (100 severe falciparum malaria (with mean age 8.63 ± 3.40 years;
61% male; 39% female), 100 uncomplicated falciparum malaria (with mean age 8.83 ± 4.20 years; 45% male; 55% female) and 100
normal healthy children controls (with mean age 10.08 ± 3.58 years; 50% male; 50% female). The malaria parasitemia was estimated
using thick blood film, while parasite count was counted using thin blood film. TNF-α levels measured using Human TNF-α ELISA
MAX™ Deluxe Sets - BioLegend, Inc. The data were analyzed using SPSS software (V 20.0) and Statdisk software (V 13.0). The average
of TNF-α levels were markedly elevated in serum of severe and uncomplicated falciparum malaria (200.98 ± 92.77 and 112.42 ± 35.52
pg/ml respectively) versus normal healthy children controls (38.81 ± 22.23 pg/ml), giving statistically highly significant difference (P
value = 0.000). Furthermore, the levels of TNF-α in malaria parasitemia had statistically highly significant differences and significant
positive correlation (P value = 0.000 for both). On the other hand, the levels of TNF-α had significant positive correlation within
parasite count (P value = 0.000). This study showed that TNF-α levels had association with falciparum malaria severity, and level of
parasitemia. The results obtained in this study will help clinicians to diagnose and to improve the management of severe malaria cases.

published in International Journal of Academic Health and Medical Research (IJAHMR)

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