Mycetoma is a devastating neglected tropical disease, caused by various fungal and bacterial pathogens. Correct diagnosis to the species level is mandatory for proper treatment. In endemic areas, various diagnostic tests and techniques are in use to achieve that, and that includes grain culture, surgical biopsy histopathological examination, fine needle aspiration cytological (FNAC) examination and in certain centres molecular diagnosis such as PCR. In this retrospective study, the sensitivity, specificity and diagnostic accuracy of grain culture, surgical biopsy histopathological examination and FNAC to identify the mycetoma causative organisms were determined. The histopathological examination appeared to have better sensitivity and specificity. The histological examination results were correct in 714 (97.5%) out of 750 patients infected with Madurella mycetomatis, in 133 (93.6%) out of 142 patients infected with Streptomyces somaliensis, in 53 (74.6%) out of 71 patients infected with Actinomadura madurae and in 12 (75%) out of 16 patients infected with Actinomadura pelletierii. FNAC results were correct in 604 (80.5%) out of 750 patients with Madurella mycetomatis eumycetoma, in 50 (37.5%) out of 133 Streptomyces somaliensis patients, 43 (60.5%) out of 71 Actinomadura madurae patients and 11 (68.7%) out of 16 Actinomadura pelletierii. The mean time required to obtain the FNAC result was one day, and for the histopathological examinations results it was 3.5 days, and for grain it was a mean of 16 days. In conclusion, histopathological examination and FNAC are more practical techniques for rapid species identification than grain culture in many endemic regions.
Appropriate diagnosis and treatment of eumycetoma may vary significantly depending on the causative agent. To date, the most common fungus causing mycetoma worldwide is Madurella mycetomatis. This species fails to express any recognizable morphological characteristics, and reliable identification can therefore only be achieved with the application of molecular techniques. Recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP) are proposed as alternatives to phenotypic methods. Species-specific primers were developed to target the ribosomal DNA (rDNA) internal transcribed spacer (ITS) region of M. mycetomatis. Both isothermal amplification techniques showed high specificity and sufficient sensitivity to amplify fungal DNA and proved to be appropriate for detection of M. mycetomatis. Diagnostic performance of the techniques was assessed in comparison to conventional PCR using biopsy specimens from eumycetoma patients. RPA is reliable and easy to operate and has the potential to be implemented in areas where mycetoma is endemic. The techniques may be expanded to detect fungal DNA from environmental samples. Madurella mycetomatis is the most common causative agent of human eumycetoma worldwide, as evidenced in a recent review and meta-analysis on the disease (1). This highly morbid and destructive infection mainly affects individuals of low socioeconomic status in arid climate zones in the tropics (2). Although mycetoma was introduced into the medical literature in the 18th century, it still remains neglected and it has recently been recognized by the World Health Organization as a neglected tropical condition (2-4). The prevalence of the infection in villages in Sudan where the condition is endemic is estimated to be 14 per 1,000 inhabitants (5). At the Mycetoma Research Centre, Khartoum, Sudan, the main reference center in the country, the annual incidence of new cases is Ͼ300 (6). Despite this high incidence, appropriate diagnoses and therapies for this disease are still lacking (7).Mycetoma is a subcutaneous infection that mainly affects the extremities and leads to massive disfigurement and disability (8, 9). The infection is thought to occur as a result of transcutaneous trauma, which allows insertion of the causative agent into skin and subcutaneous tissue. A distinctive and unique feature of mycetoma agents is their ability to form compact grains in infected tissue that will be discharged through the sinuses (10). Since mycetoma can be caused by both bacteria (actinomycetoma) and fungi (eumycetoma), the discharged grains are different (11). Black grains are caused by fungi, while actinomycetoma grains are white, yellow, or red (2, 10, 12). Molecular phylogeny of the eumycetoma causative agents has shown that the black-grain agents belong to entirely different fungal species, and these fungi differ in their antifungal susceptibility profiles (13,14). Therefore, precise identification of the causative agent is necessary.Current techniques for mycetoma diagnostics mostly co...
The first yellow-grain fungal mycetoma, in a 60-year-old man from Central Sudan, is reported. Morphological and phylogenetic analysis of the ribosomal small subunit (SSU), large subunit (LSU), internal transcribed spacer (ITS), β-tubulin ( BT2 ), actin ( ACT1 ), and elongation factor ( TEF1 ) genes revealed that the isolate deviated from any known agent of mycetoma; it clustered in the genus Pleurostoma (anamorph genus, Pleurostomophora ) in the order Calosphaeriales . The novel species, here named Pleurostomophora ochracea , is characterized by phenotypic features. The species proved to be highly susceptible to itraconazole, ketoconazole, posaconazole, and voriconazole, but not to fluconazole. The fungus was inhibited by caspofungin at 8 μg/ml, while no inhibition was found with 5-flucytosine (MIC > 64 μg/ml). Compared to other members of the genus Pleurostomophora , P. ochracea is slow growing, with a relatively high optimum growth temperature (36 to 37°C). This is the first case of a yellow-grain fungal mycetoma; yellow grains are otherwise of bacterial nature. Our case emphasizes that identification of mycetoma agents by the color of the grain only is not sufficient and may lead to inappropriate therapy.
Abstractobjective To investigate the genetic determinants for developing tuberculosis in Sudan. methods Case study of 232 patients with tuberculosis and 206 healthy matched controls from Sudan. In the study population, three single nucleotide polymorphisms (SNPs) in the promoter regions of CCL5 and two in the promoter region of IL-10 were genotyped using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). These five SNPs influence the expression of these genes.
Mycetoma is a progressive and destructive chronic granulomatous subcutaneous inflammatory disease caused by bacteria and fungi. The genetic determinants for susceptibility to and the development of mycetoma are unclear. Polymorphisms in genes encoding for cytokines and chemokines usually influence the efficiency of the immune response to infection and are associated with disease susceptibility and progression. Therefore, we hypothesized that polymorphisms of CC chemokine ligand 5 (CCL5) and interleukin-10 (IL-10) promoter regions might contribute to the initiation, susceptibility, and severity of eumycetoma. This case-control study included 149 mycetoma patients and 206 healthy matched controls. In the study population, three functional single nucleotide polymorphisms (SNPs) in CCL5 and two in IL-10 were genotyped using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). Significant differences in allele distribution were demonstrated for CCL5 -28 C/G (P < 0.0001), CCL5 In1.1 T/C (P < 0.0001) and IL-10 -592 A/C. Since in previous studies it was demonstrated that the genotypes obtained for CCL5 and IL-10 were connected with CCL5 and IL-10 production we measured the serum levels of CCL5 and IL-10 in mycetoma patients and healthy controls. Elevated serum levels for both CCL5 and IL-10 were found in mycetoma patients and we describe that genetic differences in CCL5 and IL-10 are associated with the development of the mycetoma granuloma.
Species of the genus Microascus are uncommon agents of human diseases despite their ubiquitous presence in the environment. In this communication, the first case of white grain eumycetoma caused by the fungus Microascus gracilis is reported. The patient was initially misdiagnosed as having actinomycetoma based on the grains morphological and cytological features and was treated with antimicrobial therapy with no clinical improvement. She underwent wide local surgical excision to improve the response to medical treatment and further grain cultural, molecular and taxonomy techniques were conducted and the diagnosis of mycetoma due to M. gracilis was established. The antifungal susceptibilities of this isolate to nine drugs were tested in vitro and they showed poor activity. Combination therapy with surgery and itraconazole led to complete recovery. A medical literature search revealed no previous report on M. gracilis as a causative agent of eumycetoma and hence we are reporting this new causative agent of human eumycetoma. Also, the difficulty in the management of this patient emphasizes the need for accurate and appropriate diagnostic tests for the identification of mycetoma-causative organisms and thus proper management.
Eumycetoma is a chronic progressive disabling and destructive inflammatory disease which is commonly caused by the fungus Madurella mycetomatis. It is characterized by the formation of multiple discharging sinuses. It is usually treated by antifungal agents but it is assumed that the therapeutic efficiency of these agents is reduced by the co-existence of Staphylococcus aureus co-infection developing in these sinuses. This prospective study was conducted to investigate the safety, efficacy and clinical outcome of combined antibiotic and antifungal therapy in eumycetoma patients with superimposed Staphylococcus aureus infection. The study enrolled 337 patients with confirmed M. mycetomatis eumycetoma and S. aureus co-infection. Patients were allocated into three groups; 142 patients received amoxicillin-clavulanic acid and ketoconazole, 93 patients received ciprofloxacin and ketoconazole and 102 patients received ketoconazole only. The study showed that, patients who received amoxicillin-clavulanic acid and ketoconazole treatment had an overall better clinical outcome compared to those who had combined ciprofloxacin and ketoconazole or to those who received ketoconazole only. In this study, 60.6% of the combined amoxicillin-clavulanic acid/ketoconazole group showed complete or partial clinical response to treatment compared to 30.1% in the ciprofloxacin/ketoconazole group and 36.3% in the ketoconazole only group. The study also showed that 64.5% of the patients in the ciprofloxacin/ketoconazole group and 59.8% in the ketoconazole only group had progressive disease and poor outcome. This study showed that the combination of amoxicillin-clavulanic acid and ketoconazole treatment is safe and offers good clinical outcome and it is therefore recommended to treat eumycetoma patients with Staphylococcus aureus co-infection.
Objectives: Infection with the bacteria Helicobacter pylori has been classified as class one carcinogen associated with increasing susceptibility of gastritis and gastric carcinoma. This study is aiming at investigating the prevalence of H. pylori among colon polyps and colon cancer patients. A descriptive cross-sectional hospital-based study was conducted between February and June 2017. Sixty-nine formalin-fixed paraffin blocks collected from colon polyps and colon cancer patients to detect H. pylori using immunohistochemistry technique. Results: Of the 69 patients included in the study, 39 (56.5%) males and 30 (43.5%) were females, their age ranged from 21 to 80 years with a mean age of 47.1 ± 19.7. Of the 69 colon polyps and colon cancer patients, 44 (63.8%) were diagnosed as adenocarcinoma, 10 (14.5%) colitis, 15 (21.7%) juvenile polyposis syndrome. The results of immunohistochemistry technique showed the presence of 16 (23.2%) positive patients for H. pylori infection. Of these 16, 13 (81.3%) patients were diagnosed with adenocarcinoma and 3 (18.7%) patients were diagnosed with juvenile polyps. The results of H. pylori detection among the different colon polyps and colon cancer patients were showing a statistically significant association for H. pylori infection and adenocarcinoma, P value 0.028.
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