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1.
Saudi Med J ; 44(2): 194-201, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36773975

RESUMO

OBJECTIVES: To assess the different side effects of COVID-19 vaccines at different scenarios in Saudi Arabia. METHODS: This cross-sectional study sought to investigate the side effects of COVID-19 vaccines through an online survey of 2,718 participants in Saudi Arabia. RESULTS: People can manage their expectations about vaccine side effects and deal with symptoms better by knowing beforehand that they are likely to experience mild side effects for a short period, symptoms that are manifested regardless of age, and infection before or after vaccination. There are certain uncommon side effects that affect more people who got infected, and not before vaccination; there are side effects that disproportionately impact women, and also the side effects that wane after the second dose. CONCLUSION: These findings can assist in evaluating the concerns regarding vaccine acceptance. The public should be made aware that they are likely to experience at least one side effect, with temporary post-injection inflammation, musculoskeletal pain, fever, and headache as the most commonly reported side effects across the board. However, the common symptoms are mild to moderate, and the side effects last for a short period for most people.


Assuntos
Vacinas contra COVID-19 , COVID-19 , Feminino , Humanos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Vacinas contra COVID-19/efeitos adversos , Estudos Transversais , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Arábia Saudita/epidemiologia
2.
Clin Lab ; 68(11)2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36378000

RESUMO

BACKGROUND: This study evaluates the seroprevalence of immunoglobulin M (IgM) and G (IgG) antibodies against SARS-CoV-2 after two doses of Pfizer-BioNTech COVID-19 vaccination from women with breast cancer in Jazan city Kingdom of Saudi Arabia, antibody detections were performed one month and three months after the administration of the second dose. METHODS: Overall, 103 breast cancer patients were included. Individuals who had had two doses of Pfizer-BioNTech vaccine, patients who were earlier diagnosed with COVID-19 infection, had not finalized immunization plan, or who received the second dose recently were excluded from the study. The antibodies detection test was run according to the manufacturer's directions of Viva Diag™ SARS-CoV-2 IgM/IgG Rapid Test (COVID-19 IgM/IgG Rapid Test). RESULTS: This study included 62 (60.2%) and 41 (39.8%) patients with invasive ductal carcinoma and invasive lobular carcinoma, respectively. The age, median and interquartile range (IQR) was 54.0 (26) years. Regarding reactivity of antibodies, after one month IgM antibody showed 64 (62.1%) positive and 39 (37.9%) negative while IgG antibody showed positive results in all patients. After three months IgM antibody showed 44 (42.7%) positive and 59 (57.3%) negative, while IgG showed 87 (84.5%) positive and 16 (15.5%) negative. There were significant differences in the IgM and IgG seropositivity. There were 19.3% patients with ductal carcinoma who were positive and then turned negative versus 17.7% who were positive and then turned negative, respectively (p < 0.001). There were significant differences in IgM antibody positivity among different age groups. CONCLUSIONS: Our results recommend the importance of screening for an antibody response for breast cancer patient after immunization in order to reveal persons who need early and late extra enhancing vaccine dose. Upcoming studies recommended to estimate different methods that raise cancer patients' immune response.


Assuntos
Neoplasias da Mama , COVID-19 , Carcinoma Ductal , Humanos , Feminino , Pessoa de Meia-Idade , SARS-CoV-2 , Imunoglobulina M , Estudos Soroepidemiológicos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Vacina BNT162 , Vacinas contra COVID-19 , Anticorpos Antivirais , Imunoglobulina G
3.
Clin Lab ; 68(6)2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35704713

RESUMO

BACKGROUND: The aim of this study was to evaluate renal function by urinalysis in COVID-19 patients following the administration of vancomycin. METHODS: A retrospective observational study was performed between October 2020 and January 2021, during which time patients were hospitalized in the Prince Mohammed Bin Abdulaziz Hospital in Riyadh, Saudi Arabia. The patients were free of kidney disease. Urinalysis was performed by an automated laboratory system, and the collected results were based upon age, gender, diabetic status, whether the patients had received vancomycin, the mortality rate, and the urinalysis panel including coinfection by bacteria and yeast. RESULTS: A total of 227 patients were included in this study, 147 (64.75%) of whom were male and 80 (35.25%) of whom were female; 54.63% were diabetic, 11.89% were prediabetic, and 33.48% were non-diabetic patients. Proteinuria, hematuria, glycosuria, coinfection, and ketonuria were detected among all participants within the study group, specifically among diabetic patients. The mortality rate was 16.2% among the study group; 6.6% had re-ceived vancomycin, and 9.6% had not received vancomycin. No significant correlation was found between nephrotoxicity and abnormalities in the urine and the mortality rate among members of our study group. CONCLUSIONS: Proteinuria, hematuria, glycosuria, ketonuria, and coinfection were common among members of our study group, especially in the diabetic group. Urinalysis abnormalities were less frequent in the vancomycin group than in the others, except the prediabetic group. No correlation between mortality and vancomycin was identified.


Assuntos
COVID-19 , Coinfecção , Glicosúria , Cetose , Estado Pré-Diabético , Feminino , Hematúria , Humanos , Rim/fisiologia , Masculino , Proteinúria , Estudos Retrospectivos , Urinálise , Vancomicina/efeitos adversos
4.
Pak J Biol Sci ; 24(11): 1169-1174, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34842389

RESUMO

<b>Background and Objective:</b> In recent years, respiratory tract viral infections have caused many pandemics that impact the whole world. To investigate the seropositivity of <i>Toxoplasma gondii</i>, rubella, CMV, HSV-1 and group A <i>Streptococcus</i> in recovered COVID-19 patients and correlate these findings with vitamin D levels. <b>Materials and Methods:</b> A total of 417 COVID-19 patients with diarrhoea were enrolled in this study. Vitamin D and seroprevalence for <i>Toxoplasma gondii</i>, rubella, CMV, HSV-1 and group A <i>Streptococcus</i> were evaluated and correlated. <b>Results:</b> It was found that recent infection in COVID-19 patients with HSV-1, rubella, <i>Toxoplasma</i> and CMV, respectively. IgG was detected indicating the development of adaptive immunity with all microbes. <b>Conclusion:</b> Current study detected a correlation between vitamin D levels and HSV-1 and no correlation between this infection and vitamin D deficiency with the other microbes.


Assuntos
Teste Sorológico para COVID-19 , COVID-19/diagnóstico , Calcifediol/sangue , Herpes Simples/diagnóstico , Herpesvirus Humano 1/imunologia , Imunoglobulina G/sangue , Deficiência de Vitamina D/diagnóstico , Imunidade Adaptativa , Adulto , Biomarcadores/sangue , COVID-19/sangue , COVID-19/epidemiologia , COVID-19/imunologia , Citomegalovirus/imunologia , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/epidemiologia , Infecções por Citomegalovirus/imunologia , Feminino , Herpes Simples/sangue , Herpes Simples/epidemiologia , Herpes Simples/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prevalência , Rubéola (Sarampo Alemão)/sangue , Rubéola (Sarampo Alemão)/diagnóstico , Rubéola (Sarampo Alemão)/epidemiologia , Rubéola (Sarampo Alemão)/imunologia , Vírus da Rubéola/imunologia , Arábia Saudita/epidemiologia , Estudos Soroepidemiológicos , Infecções Estreptocócicas/sangue , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/imunologia , Streptococcus/imunologia , Toxoplasma/imunologia , Toxoplasmose/sangue , Toxoplasmose/diagnóstico , Toxoplasmose/epidemiologia , Toxoplasmose/imunologia , Deficiência de Vitamina D/sangue , Deficiência de Vitamina D/epidemiologia
5.
Biomed Res Int ; 2021: 1622270, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34409099

RESUMO

This study investigates the antioxidant activities of lipid, protein, and carbohydrate extracts from the marine mollusk Perna canaliculus. Lipids were extracted using acetone, which was followed by protein extraction using the broad-spectrum enzyme Alcalase and then carbohydrate extraction using cetylpyridinium chloride. Eighty white BALB/c mice were divided into eight groups according to the administered extracts. Groups 1 and 5 were the control and toxin control groups, respectively. Groups 2, 3, and 4 were administered lipid, protein, and carbohydrate extracts, respectively. The other groups were administered P. canaliculus extracts as well as gentamicin and acetaminophen, known as ethanolic extracts, derived from Nerium oleander to induce oxidation stress. All groups showed significant improvements in body weight (p < 0.05). The lipid extract group showed a significant decrease in low-density lipoprotein cholesterol (p < 0.05) and a significant increase in high-density lipoprotein cholesterol (p < 0.05). After the toxin injection, all groups treated with P. canaliculus extracts showed increased antioxidant effects on hepatocytes (p < 0.05). The lipid extracts induced antioxidant effects to protect the kidney by increasing lipid peroxidation (p < 0.05) and catalase activities (p < 0.05). Also, protein extracts showed antioxidant effects by increasing glutathione and catalase levels significantly (p < 0.005). In conclusion, P. canaliculus extracts, especially lipids and proteins, have potent antioxidant activities that protect vital organs from oxidation stress.


Assuntos
Antioxidantes/administração & dosagem , Carboidratos/administração & dosagem , Lipídeos/administração & dosagem , Perna (Organismo)/química , Proteínas/administração & dosagem , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Produtos Biológicos/isolamento & purificação , Carboidratos/isolamento & purificação , Carboidratos/farmacologia , Catalase/metabolismo , Etanol/administração & dosagem , Etanol/farmacologia , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Lipídeos/isolamento & purificação , Lipídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Nerium/química , Estresse Oxidativo/efeitos dos fármacos , Proteínas/isolamento & purificação , Proteínas/farmacologia
6.
J Food Biochem ; 45(9): e13865, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34263474

RESUMO

In the past decade, the use of marine mussels as seafood is being more popular. They considered being a rich source of various nutritional bioactive compounds that aroused the scientific community's interest. This study investigated the antioxidant and the antithrombotic consequences on Sprague-Dawley male rats after adding dried New Zealand mussel Perna canaliculus in their diet. The biochemical, hematological and histopathological changes were also observed. Forty rats were divided into four groups according to the amount of dried mussels in their diet, in addition to a control group that consumed a basal diet only. Group 1 consumed 25% dried mussels in its basal diet; Group 2 consumed 35% dried mussels in its basal diet, and Group 3 was consumed 45% dried mussels in its basal diet. The biochemical parameters showed improvements in liver function. Interestingly, the lipid profile decreased, especially the low-density lipoprotein cholesterol (LDL-C) levels which were reduced significantly in Group 3 (p < .01). These observations were accompanied by a decrease in iron levels significantly as the amount of dried mussels increased (p < .01). Furthermore, the noticed changes in the hematological profile prove that there is an increase in antithrombotic activity. Dried mussels had potent antioxidant effects in the liver as shown by increased lipid peroxide (p < .05), reduced glutathione (p < .05), and glutathione peroxidase (GSH-Px; p < .05). Additionally, antioxidant activity in the kidney was shown to increase through GSH-Px activity (p < .01). In conclusion, these results indicate that consuming dried mussels resulted in improved biochemical and antioxidants activities and could be used as an antithrombotic agent.


Assuntos
Perna (Organismo) , Animais , Antioxidantes/farmacologia , Fibrinolíticos/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley , Alimentos Marinhos
7.
Pak J Biol Sci ; 23(11): 1369-1373, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33274863

RESUMO

BACKGROUND AND OBJECTIVES: Microcystin-LR has a toxic effect on several organs causing the destruction and fibrosis of these organs. This study was done to evaluate the antioxidant activity of silymarin on some oxidative stress parameters on heart and lung injuries induced by microcystin-LR in mice. MATERIALS AND METHODS: Total 72 Balb/c male mice aged between five to seven weeks were grouped into 6; Group 1 contained twelve mice which were assigned as the healthy control group (C). Two microcystin-LR control groups assigned M6 and M12 contain 12 mice each. A fourth group contains twelve mice called the silymarin control group(S). The fifth and sixth groups contain twenty-four mice assigned as microcystin-LR silymarin groups SM6 and SM12. A blood sample was collected for estimation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The homogenates of heart and lungs were extracted for estimation of protein carbonyl content (CC), lipid peroxidation products (LPO), reduced glutathione (GSH), methylglyoxal (MG) and activity of protein phosphatase I (PPI). RESULTS: Significant statistical differences in both ALT and AST were observed between all groups (P>0.01). In cardiac homogenate, a significant statistical difference was observed in PPI, LPO and CC between all groups (P<0.05). Furthermore, methylglyoxal showed a significant statistical difference between all groups (p<0.01). CONCLUSION: The findings suggested a potential therapeutic role of using silymarin as an antioxidant agent against cardiac and pulmonary injuries induced by MC-LR.


Assuntos
Antioxidantes/farmacologia , Cardiopatias/tratamento farmacológico , Pulmão/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fibrose Pulmonar/tratamento farmacológico , Silimarina/farmacologia , Animais , Biomarcadores/metabolismo , Cardiotoxicidade , Modelos Animais de Doenças , Glutationa/metabolismo , Cardiopatias/induzido quimicamente , Cardiopatias/metabolismo , Cardiopatias/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Toxinas Marinhas , Camundongos Endogâmicos BALB C , Microcistinas , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Carbonilação Proteica/efeitos dos fármacos , Proteína Fosfatase 1/metabolismo , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Aldeído Pirúvico/metabolismo
8.
Pak J Biol Sci ; 23(12): 1676-1680, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33274902

RESUMO

BACKGROUND AND OBJECTIVE: Commiphora gileadensis is a plant in the Burseraceae family that grows in the western area of Saudi Arabia. Traditionally, it is used in the treatment of some superficial infections. MATERIALS AND METHODS: The methanolic extract of Commiphora gileadensis isolated from its leaves and branches. The in vitro study was conducted to determine the effect of this extract on Methicillin-Resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa using an agar diffusion and Minimum inhibitory concentration (MIC) methods. The in vivo study was conducted through two different methods. The first method, 20 male Balb c-1 mice were used for the determination of Commiphora gileadensis methanolic extract toxicity (LD50). In the second method, 40 male mice were used and were put into four groups. The first and second groups were injected subcutaneously with 108 CFU of MRSA 1 mL-1, while the third and fourth groups were injected with 108 CFU of Pseudomonas aeruginosa 1 mL-1. The comparison between groups was done by using a t-test (p<0.05). RESULTS: The methanolic extract of Commiphora gileadensis had a greater sensitivity zone on MRSA and Pseudomonas aeruginosa, 7 and 3 mm respectively. The MIC of the extract was 1/8 and 1/2 for MRSA and Pseudomonas aeruginosa respectively. The in vivo study showed that the extract was non-toxic, it also showed that the extract decreased the mortality of mice induced by MRSA injection significantly (p<0.05) While insignificantly with Pseudomonas aeruginosa. CONCLUSION: The total Commiphora gileadensis methanolic extract had an antibacterial effect on MRSA and Pseudomonas aeruginosa. This extract was non-toxic for the mice.


Assuntos
Antibacterianos/farmacologia , Commiphora , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Extratos Vegetais/farmacologia , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/efeitos dos fármacos , Infecções Estafilocócicas/tratamento farmacológico , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/toxicidade , Commiphora/química , Commiphora/toxicidade , Modelos Animais de Doenças , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Dose Letal Mediana , Masculino , Metanol/química , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Camundongos Endogâmicos BALB C , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/crescimento & desenvolvimento , Solventes/química , Infecções Estafilocócicas/microbiologia
9.
Front Cell Dev Biol ; 8: 444, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32637407

RESUMO

Sphingosine-1-phosphate (S1P) is a pleiotropic sphingolipid derived by the phosphorylation of sphingosine either by sphingosine kinase 1 (SPHK1) or SPHK2. Importantly, S1P acts through five different types of G-protein coupled S1P receptors (S1PRs) in immune cells to elicit inflammation and other immunological processes by enhancing the production of various cytokines, chemokines, and growth factors. The airway inflammation in asthma and other respiratory diseases is augmented by the activation of immune cells and the induction of T-helper cell type 2 (Th2)-associated cytokines and chemokines. Therefore, studying the S1P mediated signaling in airway inflammation is crucial to formulate effective treatment and management strategies for asthma and other respiratory diseases. The central aim of this study is to characterize the molecular targets induced through the S1P/S1PR axis and dissect the therapeutic importance of this key axis in asthma, airway inflammation, and other related respiratory diseases. To achieve this, we have adopted both high throughput next-generation knowledge discovery platforms such as SwissTargetPrediction, WebGestalt, Open Targets Platform, and Ingenuity Pathway Analysis (Qiagen, United States) to delineate the molecular targets of S1P and further validated the upstream regulators of S1P signaling using cutting edge multiple analyte profiling (xMAP) technology (Luminex Corporation, United States) to define the importance of S1P signaling in asthma and other respiratory diseases in humans.

10.
Saudi Med J ; 40(6): 555-559, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31219489

RESUMO

OBJECTIVES: To determine human papillomavirus (HPV) infection, cervical cancer, and vaccine awareness among the Saudi population. METHODS: This cross-sectional study of a convenience sample comprising 1033 participants (males and females) from different parts of Kingdom of Saudi Arabia was conducted between August 2018 and January 2019 using a web-based questionnaire. This self-administrated questionnaire was distributed to all participants. Collected data included age groups, cervical cancer, Papanicolaou (Pap) smear, and HPV vaccine awareness. RESULTS: The response rate was 95%. Approximately 50% of the participants were 15-22 years old, less than 3% were more than 46 years old, and less than 10% had heard of HPV. Awareness and previous knowledge of the Pap smear as a screening tool was variable with male (5.9%) and female (27.9%) participants, having knowledge of the test. There were no statistically significant differences (p more than 0.05) between males and females in their knowledge of HPV's role in cervical and penile cancers, the HPV vaccine availability in the hospital, its role in cervical cancer prevention, and suggestions that this vaccine should be provided to married and non-married women. CONCLUSION: There is a lack of knowledge and misinformation regarding cervical cancer, Pap smears, HPV, and HPV association with cervical cancer. These data can be used as a basis to formulate effective population awareness programs.


Assuntos
Conscientização , Conhecimento , Infecções por Papillomavirus/prevenção & controle , Neoplasias do Colo do Útero/prevenção & controle , Adolescente , Adulto , Fatores Etários , Estudos Transversais , Feminino , Humanos , Programas de Imunização , Masculino , Pessoa de Meia-Idade , Infecções por Papillomavirus/complicações , Vacinas contra Papillomavirus , Neoplasias Penianas/etiologia , Neoplasias Penianas/prevenção & controle , Vigilância da População , Arábia Saudita , Inquéritos e Questionários , Neoplasias do Colo do Útero/etiologia , Adulto Jovem
11.
Afr Health Sci ; 19(4): 2812-2818, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32127856

RESUMO

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a global problem. Association of multiple genes in T2DM becomes a hot point recently. This study was aimed to evaluate association of vitamin D receptor gene polymorphisms with susceptibility to T2DM. SUBJECTS AND METHODS: One hundred T2DM Saudi male patients were included in this study and one hundred healthy Saudi men were used as control. For each individual, fasting blood glucose, cholesterol, HDL-C, LDL-C, HbA1c, insulin and 25-(OH) vitamin D were measured. In addition, Apal, BsmI and TaqI genotypes were performed for each subject. Data was analyzed by SPSS version 16, using Spearman's rho and ANOVA tests. RESULTS: There was significant inverse correlation between 25-(OH) vitamin D level and T2DM (p<0.01). HbA1c was inversely correlated with 25-(OH) vitamin D level (P<0.05). Genotype study showed that tt of TaqI genotype was higher in T2DM group compared with control group (p<0.05). Moreover, tt genotype has higher HbA1c than both TT and Tt genotypes (p<0.05). CONCLUSION: An association was confirmed between TaqI genotypes and T2DM but there is no correlation between BsmI, ApaI and T2DM. In addition, HbA1c is positively correlated with tt genotype of TaqI.


Assuntos
Diabetes Mellitus Tipo 2/genética , Polimorfismo Genético , Receptores de Calcitriol/genética , Deficiência de Vitamina D/genética , Adulto , Estudos de Casos e Controles , Estudos de Associação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único
12.
Electron Physician ; 9(10): 5427-5433, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29238479

RESUMO

BACKGROUND: The prevalence of obesity and vitamin D deficiency in Saudi Arabia has increased recently. Decreased physical activity might play a role in obesity. Previous studies showed an association between low vitamin D level and its receptor polymorphism with obesity development. OBJECTIVE: To determine association of low vitamin D level and its receptor polymorphism with obesity in Saudi men. METHODS: This case control study was carried out from March 2016 through March 2017. Three hundred Saudi male students (from applied medical sciences in Taif University, Taif, Saudi Arabia) were classified according to BMI into lean, overweight and obese groups. For each individual, blood glucose, cholesterol, HDL-C, LDL-C, insulin and 25-(OH) vitamin D were measured. In addition, Apal, BsmI and TaqI genotypes were performed for each individual from March 2016 through March 2017, through computer-based search of the following databases: PubMed, Web of Science (Thomson ReutersTM). The references of the original literature and the related articles were also searched, for potential complementary studies. Data were analyzed by SPSS version 16, using Spearman's rho and ANOVA tests. RESULTS: There was significant negative association between 25-(OH) vitamin D level and obesity (p<0.01). Genotyping study showed that both bb of BsmI and tt of TaqI genotypes were higher in the obese group compared with lean group (p<0.05). Moreover, bb genotype has higher BMI and HOMA-IR than both BB and Bb; and tt genotype also has higher BMI and HOMA-IR than TT and Tt genotypes (p<0.05). CONCLUSION: Low vitamin D level and VDR BsmI and Taq1 genotypes may be a risk factor of obesity.

13.
Korean J Parasitol ; 53(2): 147-54, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25925172

RESUMO

Various constituents in clinical specimens, particularly feces, can inhibit the PCR assay and lead to false-negative results. To ensure that negative results of a diagnostic PCR assay are true, it should be properly monitored by an inhibition control. In this study, a cloning vector harboring a modified target DNA sequence (≈375 bp) was constructed to be used as a competitive internal amplification control (IAC) for a conventional PCR assay that detects ≈550 bp of the Cryptosporidium oocyst wall protein (COWP) gene sequence in human feces. Modification of the native PCR target was carried out using a new approach comprising inverse PCR and restriction digestion techniques. IAC was included in the assay, with the estimated optimum concentration of 1 fg per reaction, as duplex PCR. When applied on fecal samples spiked with variable oocysts counts, ≈2 oocysts were theoretically enough for detection. When applied on 25 Cryptosporidium-positive fecal samples of various infection intensities, both targets were clearly detected with minimal competition noticed in 2-3 samples. Importantly, both the analytical and the diagnostic sensitivities of the PCR assay were not altered with integration of IAC into the reactions. When tried on 180 randomly collected fecal samples, 159 were Cryptosporidium-negatives. Although the native target DNA was absent, the IAC amplicon was obviously detected on gel of all the Cryptosporidium-negative samples. These results imply that running of the diagnostic PCR, inspired with the previously developed DNA extraction protocol and the constructed IAC, represents a useful tool for Cryptosporidium detection in human feces.


Assuntos
Criptosporidiose/diagnóstico , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Reação em Cadeia da Polimerase/normas , Cryptosporidium/genética , Primers do DNA/genética , DNA de Protozoário/genética , Fezes/parasitologia , Humanos , Reação em Cadeia da Polimerase/métodos , Padrões de Referência
14.
Korean J Parasitol ; 52(5): 479-85, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25352695

RESUMO

Cryptosporidium diarrhea represents a relevant clinical problem in developing countries. In Al-Taif, a city of Saudi Arabia that lies at an altitude of an around 2 km above the sea level, Cryptosporidium infection seems to be undiagnosed in nearly all clinical laboratories. Furthermore, nothing was published regarding Cryptosporidium-associated diarrhea in this area. The objectives of this research were to (1) determine the Cryptosporidium prevalence among patients with diarrhea and (2) to estimate the performances of 3 different diagnostic methods. Total 180 diarrheal fecal samples, 1 sample per patient, were collected between January and August 2013. Samples were screened for Cryptosporidium with modified Zeihl Neelsen (ZN) microscopy, RIDA® Quick lateral flow (LF) immunotest, and a previously published PCR. The Cryptosporidium prevalence rate was 9.4% (17/180), 10% (18/180), and 11.6% (21/180) by microscopy, LF, and PCR test, respectively. Infection was significantly (P=0.004) predominant among children <5 years (22%) followed by children 5-9 years (11.1%). Although infection was higher in males than in females (16.2% males and 8.5% females), the difference was not statistically significant (P=0.11). Compared to PCR, the sensitivity of microscopy and the LF test were 80.9%, 85.7%, respectively. To conclude, high Cryptosporidium-associated diarrhea was found in this area especially in children ≤9 years. The PCR test showed the best performance followed by the LF test and ZN staining microscopy. The primary health care providers in Al-Taif need to be aware of and do testing for this protozoon, particularly for children seen with diarrhea.


Assuntos
Altitude , Criptosporidiose/epidemiologia , Cryptosporidium/isolamento & purificação , Diarreia/parasitologia , Adolescente , Adulto , Criança , Pré-Escolar , Diarreia/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Arábia Saudita/epidemiologia , Adulto Jovem
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