scholarly journals Conventional and molecular detection of vibrio cholerae isolated from environmental water with the prevalence of antibiotic resistance mechanisms.

2019 ◽  
Vol 10 (3) ◽  
Author(s):  
Suad A Al-Hilu ◽  
Ali M Al-Mohana ◽  
Zainab Jaber
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Molecular Detection ◽  
Public Health Problem ◽  
Environmental Water ◽  
Selective Medium ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Vibrio Cholera ◽  
Polymerase Chain

Environmental water is an important source for Vibrio cholerae, which is autochthonous to the aquatic environment, monitoring this bacterium in water is important for control of cholera. Vibrio cholerae represents an enormous public health problem around the world, especially in developing countries. One hundred samples were collected and selected. The samples were filtered and transferred to slants containing 2ml of alkaline peptone water, then subcultured on selective medium Thiosulphate Citrate Bile Salt Sucrose agar. All presumptive isolates were confirmed by using a series of biochemical tests including Oxidase test, Simmon Citrate test, DNase test, Indole test, Klingler Iron Agar (KIA) test, MacConkey agar test and motility. Confirmed Vibrio cholera strains were then screening for slide agglutination test by using commercially antisera polyvalent and monovalent O1 and O139 for determining strain serotype. The DNA extracted from pure culture and Polymerase Chain Reaction assay was used for molecular detection of Vibrio cholerae, a specific primer which designed according to ctxA gene sequences. This primer was detection and amplifying 241 base pairs of the ctxA gene. The resistance to antibiotics by Vibrio cholerae was determining by using thirteen standardized disc diffusion including Amikacin, Ceftriaxone, Ceftazidime, Gentamycin, Tetracycline, Streptomycin, Tobramycin, Cephotaxime, Nalidixic Acid, Norfloxacin, Cephalothin, Rifampicin, Cefixime. From one hundred water samples were detected, fifty-six samples were motile and positive for biochemical tests. Fifteen isolates confirmed as Vibrio cholera by Polymerase Chain Reaction (PCR) assay with primers de­signed for ctxA and 241bp band was observed. These fifteen isolates showed agglutination with polyvalent and monovalent O1 antisera, and two strains represented Ogawa from other strains that showed Inaba. The fifteen isolates exhibited an identical response to each antibiotic examined. They showed sensitive to all antibiotics except Amikacin, Streptomycin, Cefixime, Norfloxacin, Cephalothin. the aim of this study was determined the accurate method for detection of Vibrio cholerae in environmental water. In the current study, we found that the molecular method using Polymerase Chain Reaction performance using the ctxA gene-specific primers for detection of Vibrio cholerae was faster and accurate and specific.

scholarly journals Etiology of Diarrhea Requiring Hospitalization in Bangladesh by Quantitative Polymerase Chain Reaction, 2014–2018

2020 ◽  
Author(s):  
Mami Taniuchi ◽  
Kamrul Islam ◽  
Md Abu Sayeed ◽  
James A Platts-Mills ◽  
Md Taufiqul Islam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Coefficient Of Variation ◽  
Quantitative Polymerase Chain Reaction ◽  
Age Groups ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Surveillance Systems ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Diarrhea remains a major public health problem and characterization of its etiology is needed to prioritize interventions. However, most data are from single-site studies of children. We tested samples from participants of any age from 11 geographically diverse hospitals in Bangladesh to describe pathogen-specific burdens of diarrhea. Methods We utilized 2 existing diarrhea surveillance systems: a Nationwide network at 10 sentinel hospitals and at the icddr,b hospital. We tested stools from enrolled participants and nondiarrheal controls for enteropathogens using quantitative polymerase chain reaction and calculated pathogen-specific attributable fractions (AFs) of diarrhea. Results We analyzed 5516 patients with diarrhea and 735 controls. Overall, rotavirus had the highest attributable burden of diarrhea (Nationwide AF, 17.7%; 95% confidence interval [CI], 14.3–20.9%; icddr,b AF, 39.9%; 38.0–41.8%), followed by adenovirus 40/41 (Nationwide AF, 17.9%; 95% CI: 13.9–21.9%; icddr,b AF, 16.6%; 95% CI, 14.4–19.4%) and Vibrio cholerae (Nationwide AF, 10.2%; 95% CI, 9.1–11.3%; icddr,b AF, 13.3%; 95% CI: 11.9–15.1%). Rotavirus was the leading pathogen in children <5 years and was consistent across the sites (coefficient of variation = 56.3%). Adenovirus 40/41 was the second leading pathogen in both children and adults. Vibrio cholerae was the leading pathogen in individuals >5 years old, but was more geographically variable (coefficient of variation = 71.5%). Other attributable pathogens included astrovirus, norovirus, Shigella, Salmonella, ETEC, sapovirus, and typical EPEC. Conclusions Rotavirus, adenovirus 40/41, and V. cholerae were the leading etiologies of infectious diarrhea requiring hospitalization in Bangladesh. Other pathogens were important in certain age groups or sites.

scholarly journals Pathology of Fowl Paratyphoid and Molecular Detection of Its Pathogen in Layer Chickens

2021 ◽  
Vol 24 (2) ◽  
pp. 47-57
Author(s):  
J Alam ◽  
T Chakma ◽  
MS Islam ◽  
MT Islam ◽  
MAHNA ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Salmonella Enteritidis ◽  
Molecular Detection ◽  
Inflammatory Cells ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Gross Lesions ◽  
Pcr Method ◽  
Polymerase Chain

The study was aimed to ascertain the pathology of fowl paratyphoid and molecular detection of its causal agent (Salmonella spp) in chickens. Pathological and swab samples were collected from layers in Gazipur district, Bangladesh. For observing the gross and microscopic lesions of different organs necropsy and histopathology were done, and to isolate and identify the Salmonella spp, different bacteriological tests and Polymerase Chain Reaction (PCR) were performed. Swabs from 150 chickens showed 66% of salmonellosis. Gram’s staining of isolated bacteria showed pink colored rod shaped bacilli. In biochemical tests, Salmonella fermented dextrose, maltose, xylose, arabinose, dulcitol, mannitol except lactose and sucrose. Investigation of gross lesions at necropsy revealed hemorrhage and congestion in intestine, liver, spleen and ovaries. Necrotic foci were found in liver and spleen, and button like ulceration in cecal tonsils as well. Microscopic lesions included hemorrhage and focal necrosis in liver and spleen. Congestion and infiltrations of inflammatory cells were observed in small intestine. Ovary was hemorrhagic and there was infiltration of heterophils. Biochemically positive and isolated Salmonella organisms were confirmed by PCR method using invA and IE1 primers. The final results showed that a total of 91.7% Salmonella suspected cultures were confirmed as Salmonella Enteritidis. Ann. Bangladesh Agric. (2020) 24(2) : 47-57

scholarly journals Critical analysis: use of polymerase chain reaction to diagnose leprosy

2016 ◽  
Vol 52 (1) ◽  
pp. 163-169 ◽  
Author(s):  
Flaviane Granero Maltempe ◽  
Vanessa Pietrowski Baldin ◽  
Mariana Aparecida Lopes ◽  
Vera Lúcia Dias Siqueira ◽  
Regiane Bertin de Lima Scodro ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Globin Gene ◽  
Public Health Problem ◽  
Reference Laboratory ◽  
Chain Reaction ◽  
Molecular Biology Technique ◽  
Pcr Inhibitor ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Pcr Techniques

ABSTRACT Leprosy is a neglected tropical disease and an important public health problem, especially in developing countries. It is a chronic infectious disease that is caused by Mycobacterium leprae, which has a predilection for the skin and peripheral nerves. Although it has low sensitivity, slit-skin smear (SSS) remains the conventional auxiliary laboratory technique for the clinical diagnosis of leprosy. Polymerase chain reaction (PCR) is a molecular biology technique that holds promise as a simple and sensitive diagnostic tool. In the present study, the performance of two PCR methods, using different targets, PCR-LP and PCR-P, were compared with SSS with regard to leprosy diagnosis in a reference laboratory. M. leprae DNA was extracted from 106 lymph samples of 40 patients who had clinical suspicion of leprosy. The samples were subjected to both PCR techniques and SSS. Amplification of the human b-globin gene was used as PCR inhibitor control. The specificity of both PCR techniques was 100%, and sensitivity was 0.007 and 0.015 µg/ml for PCR-LP and PCR-P, respectively. No significant difference was found between either the PCR-LP or PCR-P results and SSS results (p > 0.05). Although PCR is not yet a replacement for SSS in the diagnosis of leprosy, this technique may be used as an efficient auxiliary tool for early detection of the disease, especially in endemic regions. This strategy may also be useful in cases in which SSS results are negative (e.g., in paucibacillary patients) and cases in which skin biopsy cannot be performed.

scholarly journals Chlamydia trachomatis detection in HIV infected patients using polymerase chain reaction

2013 ◽  
Vol 2 (1) ◽  
pp. 12-16
Author(s):  
A Shrestha ◽  
N Adhikari ◽  
Y Shah ◽  
P Poudel ◽  
B Acharya ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chlamydia Trachomatis ◽  
Public Health Problem ◽  
Chlamydia Infection ◽  
Hiv Positive ◽  
Chain Reaction ◽  
Hiv Patients ◽  
Important Public Health Problem ◽  
Sexually Transmitted ◽  
Polymerase Chain

Introduction: Chlamydia trachomatis is a sexually transmitted organism and causes important public health problem in the sexually active age group. Limited studies are found regarding the prevalence of C. trachomatis in Nepal. Moreover, currently there are no any study in Nepal reporting the association of chlamydia and HIV infection. This study attempts to determine the burden of chlamydia on HIV positive patients. Materials and Methods: A total of 117 HIV positive patients visiting a HIV clinic in Kathmandu, were screened for chlamydia infection. For this, urine samples were collected and analyzed using the Polymerase Chain Reaction Technique (PCR). Results: C. trachomatis was detected in 4.2% of the total 117 HIV patients. Out of positive cases 60% were males and 40% were females. However, chlamydia was found more prevalent among females (6.8%) than males (3.4%). Eighty percent of positive cases were asymptomatic. Conclusions: Although, the prevalence of chlamydia infection was found less HIV patients, most of those cases were asymptomatic. Therefore, routine checkup is recommended for all suspected cases for timely management of the disease. DOI: http://doi.dx.org/10.3126/ijim.v2i1.8003 Int J Infect Microbiol 2013;2(1):12-16

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

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