scholarly journals TRACING THE KINSHIP OF BRUCELLA SP. BY USINGA POLYMERASE CHAIN REACTION TECHNIQUE WITH A SHORT PRIMER

2020 ◽  
Vol 21 (4) ◽  
pp. 493-502
Author(s):  
Difa Widyasari ◽  
Eko Sugeng Pribadi ◽  
Fachriyan Hasmi Pasribu ◽  
Widya Septiningtyas ◽  
Jati Adiputra
Keyword(s):  
Animal Health ◽  
Small Ruminants ◽  
Chain Reaction ◽  
Cloning And Sequencing ◽  
Phylogenic Tree ◽  
Cloning Technique ◽  
Polymerase Chain ◽  
Relationship Of ◽  
Pcr Techniques ◽  
Sequencing Result

Brucellosis is a zoonotic disease that has spread throughout the world and has an important impacton both human and animal health. The four species of Brucella that cause disease in humans are Brucellaabortus, B. suis, B. melitensis and B. canis, and B. melitensis as the most pathogenic species. This Researchused 46 samples were collected from Government Small Ruminants Abattoir in Bogor Regency. Thirty twospleen samples were examined by previous research and showed a positive result when were tested withCFT and PCR techniques, but sequencing has not yet been done. Fourteen serum and spleen samples wereexamined by the similar techniques. The Research aimed to determined the genetic relationship of Brucellasp. using a PCR technique with a specific short primer to B. mellitensis. Cloning technique was appliedpreviously to five PCR positive samples before sequencing. Cloning and sequencing result of the Sample91 showed higher homology to B. melitensis and B. abortus for 127 nucleotide lengths, 97.6% -100% and99.2% -100% respectively. In the phylogenic tree, the Sample 91 was part of B melitensis sequences 1, 2,and 3 with accession numbers LT962930.1 and LT962936.1, B abortus sequences 1 and 2 with accessionnumbers CP033079.1 and B. abortus sequence 1 with accession number CP034695.1. Sample of 95, 97, 7,and 13 have lower homologies than Sample 91.

scholarly journals ANALISIS MOLEKULER FILOGENETIK DAN STRUKTUR ANTIGENIC VIRUS AVIAN INFLUENZA SUBTIPE H5N1 ISOLAT LAMPUNG TAHUN 2008-2013

2015 ◽  
Vol 9 (1) ◽  
Author(s):  
Eko Agus Srihanto ◽  
Widya Asmara ◽  
Michael Haryadi Wibowo
Keyword(s):  
Amino Acids ◽  
Avian Influenza ◽  
Animal Health ◽  
Antigenic Site ◽  
Multiple Alignment ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Phylogenic Tree ◽  
Tree Analysis ◽  
Polymerase Chain

Penelitian ini bertujuan melakukan karakterisasi molekuler antigenic site terhadap isolat virus avian influenza (AI) Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional III Lampung dari tahun 2008-2013. Amplifikasi RNA dilakukan dengan teknik reverse transcription polymerase chain reaction (RT-PCR) menggunakan 4 pasang primer referens dari Australian Animal Health Laboratory (AAHL) Geelong Australia (HA10, HA20, dan HA30) dan dilanjutkan dengan proses pengurutan. Analisis hasil pengurutan dengan menggunakan perangkat lunak MEGA versi 5.05 yang meliputi multiple alignment, deductive amino acids prediction, dan phylogenic tree analysis diperoleh hasil perbedaan genetik antar isolat Lampung dari tahun 2003-2013 ditemukan berkisar 1,1-9,1% dengan tingkat homologi mencapai 90,9-98,9%. Variasi genetik ditemukan adanya substitusi pada posisi 53 (R53K), 126 (D126E), 136 (P136), 138 (H138Q, dan H138L), 140 (R140K, R140S, dan R140N), 141 (S141P), dan 189 (K189R). Berdasarkan analisis filogenic tree isolat Lampung tahun 2008-2011 termasuk ke dalam clade 2.1.3. Analisis filogenik isolat AI tahun 2012-2013 yang menginfeksi unggas air mempunyai homologi sekitar 98,5-99,1% dibandingkan dengan isolat AI yang menginfeksi unggas air asal Jawa dan termasuk ke dalam clade 2.3.2.1.

Zebrafish embryos exposed to deltamethrin exhibit abnormalities despite induced expression of related genes (you, you-too, momo and u-boot)

2018 ◽  
Vol 35 (1) ◽  
pp. 11-19
Author(s):  
Kuder Reshma Shabnam ◽  
Dharmapuri Gangappa ◽  
Gundala Harold Philip
Keyword(s):  
Vital Role ◽  
Primary Objective ◽  
Chain Reaction ◽  
Environmental Persistence ◽  
Expression Of Genes ◽  
Pericardial Edema ◽  
Eye Pigmentation ◽  
Polymerase Chain ◽  
Relationship Of ◽  
The Relationship

Evaluation of the toxic effects of a widely used synthetic pyrethroid, deltamethrin (DM), was carried out in this study. This pesticide is preferred for pest control because of its low environmental persistence and toxicity. We investigated the expression pattern of four genes, namely, you ( you), yot ( you-too), momo ( mom) and ubo ( u-boot) during early development of zebrafish, that is, from 12 hpf to 48 hpf stages. These stages are selected as most of the important developmental aspects take place during this period. All four genes are known to play a vital role in development of notochord and somites. To understand the effect of DM on development, embryos of 4 hpf stage were exposed to two concentrations (100 and 200 µg/L) of DM, and observations were made at 12, 24 and 48 hpf stages. Our earlier studies have shown phenotypic abnormalities such as notochord bending, tail deformation, yolk sac and pericardial edema, lightening of body and eye pigmentation and interfered in somite patterning, during these stages of development. Understanding the relationship of phenotypic abnormalities with these four genes has been our primary objective. These four genes were analyzed by Reverse transcription (RT)-polymerase chain reaction and intensity of the bands has shown induction in their expression after exposure to the toxicant. In spite of the expression of genes, it was noticed that DM caused abnormalities. It can be said from the results that translational pathway could have been affected.

scholarly journals Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

2012 ◽  
Vol 3 (1) ◽  
pp. 13
Author(s):  
Aline T.A. Chagas ◽  
Michelle D. Oliveira ◽  
Jose M.S. Mezencio ◽  
Eduardo A.M. Silva ◽  
Leandro L. Oliveira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Epidemiological Surveillance ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Denv Serotypes ◽  
Polymerase Chain ◽  
Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

scholarly journals Isolation ofMycobacterium aviumsubspeciesparatuberculosisfrom non-ruminant wildlife living in the sheds and on the pastures of Greek sheep and goats

2007 ◽  
Vol 136 (5) ◽  
pp. 644-652 ◽  
Author(s):  
M. FLOROU ◽  
L. LEONTIDES ◽  
P. KOSTOULAS ◽  
C. BILLINIS ◽  
M. SOFIA ◽  
...  
Keyword(s):  
Type Strain ◽  
Insertion Sequence ◽  
Small Ruminants ◽  
Dairy Sheep ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Sheep And Goats ◽  
Faecal Samples ◽  
Polymerase Chain ◽  
Type Strains

SUMMARYThis study aimed to: (1) investigate whether non-ruminant wildlife interfacing with dairy sheep and goats of four Greek flocks endemically infected withMycobacterium aviumsubspeciesparatuberculosis(MAP) harboured MAP and (2) genetically compare the strains isolated from the wildlife to those isolated from the small ruminants of these flocks. We cultured and screened, by polymerase chain reaction (PCR), pooled-tissue samples from 327 wild animals of 11 species for the MAP-specific IS900insertion sequence. We also cultured faecal samples from 100 sheep or goats from each of the four flocks. MAP was detected in samples from 11 sheep, 12 goats, two mice, two rats, a hare and a fox. Only one rat had histopathological findings. Genetic typing categorized 21 isolates as cattle-type strains and two, from a house mouse and a goat respectively, as sheep-type strains; this is the first report of a rodent harbouring a sheep-type strain. The MAP types that were most frequently isolated amongst the sheep and goats of each flock were also the ones isolated from sympatric rodents; those isolated from the fox and hare also belonged to the predominant ruminant strains.

scholarly journals A serological and molecular investigation of American cutaneous leishmaniasis in dogs, three years after an outbreak in the Northwest of Paraná State, Brazil

2009 ◽  
Vol 25 (1) ◽  
pp. 97-104 ◽  
Author(s):  
Gustavo Kiyoshi Massunari ◽  
Evandra Maria Voltarelli ◽  
Demilson Rodrigues dos Santos ◽  
Ademar Rodrigues dos Santos ◽  
Luiz Paschoal Poiani ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cutaneous Leishmaniasis ◽  
Control Measures ◽  
Transmission Control ◽  
Chain Reaction ◽  
American Cutaneous Leishmaniasis ◽  
Molecular Investigation ◽  
Polymerase Chain ◽  
Pcr Techniques

Classic and molecular (polymerase chain reaction - PCR) techniques were used to diagnose American cutaneous leishmaniasis in 149 dogs from an area in the northwest of Paraná State, Brazil, where an American cutaneous leishmaniasis outbreak occurred in 2002. The results were compared to a set of previously obtained results. Twenty-five dogs had positive indirect immunofluorescence (IIF) (titers > 40), including two animals with suggestive lesions. The percentage of dogs with positive IIF was similar to that found in a previous study. The cultures of the lesion, blood and bone marrow were negative for Leishmania. A direct search for the parasite in the lesions proved negative, although PCR tests were positive. The PCR did not detect the DNA of Leishmania (Viannia) in the blood, even for those that had positive PCR in a previous study. The follow up of the 27 dogs showed that the majority of them had maintained the same levels of antibodies that had been detected previously. There was a reduction in the number of dogs with lesions, probably due to the transmission control measures that were adopted after the outbreak.

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.

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