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 Comparative Study of Microscopy and Polymerase Chain Reaction for the Diagnosis of Suspected Visceral Leishmaniasis Patients in Nepal

2014 ◽  
Vol 11 (1) ◽  
pp. 14-17 ◽  
Author(s):  
K Pandey ◽  
AK Mallik ◽  
S Pyakurel ◽  
SB Pun ◽  
BD Pandey
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Polymerase Chain Reaction Amplification ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Endemic Areas

Background Visceral leishmaniasis is potentially fatal protozoan diseases caused by Leishmania donovani. Nepal is an endemic region in which visceral leishmaniasis causes a major public health problem in the lowland areas that border the endemic areas of Bihar state in India. Accurate diagnosis to inform treatment is a first step in achieving the goal of visceral leishmaniasis elimination from South East Asian regions by 2020. Objective The objective of the present study was to compare between the Microcopy and polymerase chain reaction for diagnosis of visceral leishmaniasis. Methods In the present study, 236 bone marrow aspirations were collected from suspected visceral leishmaniasis patients in Janakpur Zonal Hospital, Dhanusa district, Terai region of Nepal in between 2003-2007. We evaluated bone marrow samples by microscopic examination with subsequent testing of the same sample by polymerase chain reaction and sequence analysis. Results Giemsa’s solution stained bone marrow slides stored for over five years were used for polymerase chain reaction amplification. The result showed that 71% were polymerase chain reaction positive and 56% were microscopic positive. Out of 104 microscopic negative bone marrow samples, 15% of samples were positive by polymerase chain reaction. Conclusion Polymerase chain reaction could make a very good option for diagnosis by using less or non-invasive material from visceral leishmaniasis patients in endemic areas of Nepal. DOI: http://dx.doi.org/10.3126/kumj.v11i1.11016 Kathmandu University Medical Journal Vol.11(1) 2013: 14-17

scholarly journals POLYMERASE CHAIN REACTION DETECTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM

2003 ◽  
Vol 42 (146) ◽  
pp. 65-70 ◽  
Author(s):  
Kun Young Sohn ◽  
S Shrestha ◽  
A Khagi ◽  
S S Malla ◽  
B M Pokharel ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Positive Result ◽  
Clinical Symptoms ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sensitivity Specificity ◽  
Culture Positive

ABSTRACTTuberculosis has remained to be a major public health problem in Nepal. The risk of spread of infection andemergence of drug-resistant strain has created the need for a rapid, sensitive and specific diagnostic test.In addition, clinically suspicious cases that do not give positive result in conventional laboratory test needmore sensitive test for diagnosis.In order to evaluate the possibility of incorporation of Polymerase Chain Reaction (PCR) in the diagnosis oftuberculosis, we performed a comparative study of PCR to detect Mycobacterium tuberculosis in sputumspecimens, against Ziehl-Neelsen (Z-N) stain and culture as a standard method.A total of 103 specimens were subjected to Z-N staining, culture and PCR for detecting Mycobacteriumtuberculosis. Of these, 19 were positive by Z-N stain, 26 by PCR and 25 by culture. Four stain negativespecimens showed positive result in both culture and PCR. Two specimens of stain and culture positive werePCR negative. Five specimens showed positive result only with PCR. Two culture positive specimens gavenegative results by both Z-N stain and PCR. Sensitivity, specificity, positive predictive value and negativepredictive value of PCR which were 84%, 93.5%, 80.8% and 94.9% respectively.This study showed that there is no need for PCR test for the smear positive cases. However, PCR could be apossible diagnostic tool for the confirmation of the smear negative cases that show clinical symptoms of TB.Key Words: Mycobacterium tuberculosis, Z-N stain, PCR, sensitivity, specificity.

scholarly journals Cutaneous Leishmaniasis in El-Madinna Manowra region, Saudi Arabia in 2012.

2013 ◽  
Vol 12 (3) ◽  
pp. 325-330 ◽  
Author(s):  
Nazar M Abdalla ◽  
Abdelgani M Abdelgani ◽  
Amani A Osman ◽  
Mohamed A Sarhan
Keyword(s):  
Polymerase Chain Reaction ◽  
Saudi Arabia ◽  
Cutaneous Leishmaniasis ◽  
Medical Science ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Chain Reaction ◽  
Single Male ◽  
Male Adult ◽  
Polymerase Chain

Objective: Leishmaniasis is a parasitic disease causing major public health problem in form of visceral and cutaneous types. The cutanoue leishmaniasis is caused by L. tropica, in low-land areas without reservoir; Arthroponatic leishmaniasis (ACL),  Zoonotic Cutaneous Leishmaniasis ( ZCL), in high-land. This case report involved; 25 years old Egyptian active young single male adult, stayed in Utama (75 Km far from El-Madina Manowra on the road to Makkah). He presented with three skin lesions on his arms occurred within the last 1-3 months. on examination revealed; volcano- like indurated ulcers which clinically suspected as leishmania lesions .Materials and Methods: Laboratory investigations were involved; skin smear using Giemsa stain, Leishmanin test (LST),  polymerase chain reaction (PCR), sequencing and phylogenitic analysis BLAST (NCBI).Results: Microscopy  positive LDB (leishmanin donovani bodies), Leishmanin test (LST) was negative. PCR  positive L. major . Sequence alignment were 100% with nine Iranian isolates and one Tunisian isolate. After one month of treatment with Pentostam (Sodium stibogluconate) local injections at the site of lesions the lesion progressed from ulcer to scar.Conclusion: L. major is a major species causing cutaneous leishmaniasis in Al-Medina Manowra region in Saudi Arabia. The usage of the polymerase chain reaction (PCR) is a useful diagnostic tool and help to identify the causative species.Bangladesh Journal of Medical Science Vol. 12 No. 03 July ’13 Page 325-330 DOI: http://dx.doi.org/10.3329/bjms.v12i3.13189 

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.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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