scholarly journals Prevalence and Molecular Epidemiological Data onDirofilaria immitisin Dogs from Northeastern States of India

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Sonjoy Kumar Borthakur ◽  
Dilip Kumar Deka ◽  
Saidul Islam ◽  
Dilip Kumar Sarma ◽  
Prabhat Chandra Sarmah
Keyword(s):  
Epidemiological Data ◽  
Elisa Test ◽  
Molecular Techniques ◽  
Diagnostic Methods ◽  
Molecular Tools ◽  
Stray Dogs ◽  
Concentration Technique ◽  
Working Dogs ◽  
Polymerase Chain

The aim of the present study was to determine the prevalence ofDirofilaria immitisin stray, pet, and working dogs (n=413, 266, and 103, resp.) from Guwahati (Assam) and Aizawl (Mizoram), areas located in two Northeastern States of India. Diagnostic methods applied were microscopy (wet film and Knott’s concentration technique), immunological test (Ag ELISA by SNAP 4Dx ELISA kit), and molecular tools (polymerase chain reaction and sequencing), which evidenced 11.38, 18.03, and 13.93% of positive animals, respectively. No significant differences were observed by area (18.23% versus 17.68%) nor by sex (18.1% versus 17.9%), whereas stray dogs proved more infected than other groups (P<0.05). ELISA test evidenced an overall 22.69% of occult infections, mainly in working dogs (60%), and molecular techniques detectedDirofilaria (Nochtiella) repensin 4 stray dogs from Guwahati. Characterization ofD. immitisisolates for ITS-2 region showed close identity with South Asian isolates.

scholarly journals Potential Application of PCR Based Molecular Methods in Fish Pathogen Identification: A Review

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Md. Ali Arman Ador ◽  
Md. Shameul Haque ◽  
Sulav Indra Paul ◽  
Jui Chakma ◽  
Rakib Ehsan ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Techniques ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Resistant Bacteria ◽  
Fish Disease ◽  
Fish Pathogen ◽  
Pathogen Identification ◽  
Chain Reaction ◽  
Polymerase Chain

Molecular biology developments have led to fast growth in new methods for fish disease diagnosis. Molecular diagnostic methods are rapid and more specific, more sensitive than the culture of pathogens, serology, histology, and biochemical methods which are traditionally utilized to identify causative agent fish disease. Molecular diagnostic methods are valuable for detecting specific pathogens that are difficult to culture in vitro or require a long cultivation period and it significantly more rapid in providing results compared to culture. It enables earlier informed decision-making and rapid diagnosis of bacteremia, particularly for low levels of bacteria in specimens. Molecular techniques which have the major significance are mainly PCR-based molecular diagnostic methods including Polymerase Chain Reaction (PCR), Real-Time Polymerase Chain Reaction (RT-PCR), Multiplex Polymerase Chain Reaction (multiplexPCR), and Random Amplified Polymorphic DNA (RAPD). These have been increasingly utilized to diagnose fish disease for the last recent years. Molecular diagnostic methods can detect pathogens from asymptomatic fish, so disease outbreaks could be prevented. As a consequence, antibiotic treatment can be reduced and the development of antibiotic-resistant bacteria can be eliminated. In this review paper, we attempt to summarize the potentiality of PCR-based molecular diagnostic methods and their application in fish pathogen identification.

Comparative Analysis of the Diagnostic Methods Used in Detecting River Blindness in Selected Endemic Areas of Imo State, Nigeria

2021 ◽  
Vol 6 (3) ◽  
pp. 106-115
Author(s):  
Daniel-Nwosu, E.I. ◽  
Esenwah, E.C. ◽  
Timothy, C.O.
Keyword(s):  
Polymerase Chain Reaction ◽  
Patch Test ◽  
Elisa Test ◽  
Diagnostic Methods ◽  
Onchocerca Volvulus ◽  
River Blindness ◽  
Chain Reaction ◽  
Imo State ◽  
Polymerase Chain ◽  
Endemic Areas

Onchocerciasis also known as river blindness is a chronic parasitic disease caused by the filarial worm Onchocerca volvulus. This study was a cross sectional experimental study carried out to compare the diagnostic methods used in detecting river blindness in selected endemic areas of Imo state, Nigeria. The multistage sampling technique was adopted to select samples for the study. All subjects used for this study gave an informed consent to be part of the study. Bloodless skin snips were collected from the center of the nodule or other parts of the body with the assistance of a laboratory scientist and taken to the laboratory for analysis. A total of four hundred inhabitants of the studied communities (Umulolo, Amuro, Umuna, Umunumo, Onicha, Nzerem, Umuneke and Umulewe) were examined. Out of these, the number infected by onchocerca volvulus based on Skin-Snip Microscopy, Polymerase Chain Reaction (PCR), Mazzotti test, Dietylcarbamazine (DEC) patch test and Enzyme linked Immunosorbent Assay (ELISA) test were 59, 197, 50, 107, 201 respectively. SPSS analysis using the one way ANOVA showed a significance difference (P< 0.05) in the sensitivity of the PCR, Skin Snip Microscopy, Mazzotti, DEC Patch test and ELISA used for detecting Onchocerca volvulus in all the study areas. In conclusion, the diagnostic screening efficiency of ELISA and PCR were observed to be higher than that of the other diagnostic methods analyzed. It was recommended that further evidence-based, comparative research studies on current and conventional diagnostic methods should be done to ascertain reliability, reproducibility, sensitivity and accuracy of methods used for detecting River Blindness. Keywords: River Blindness, Onchocerciasis, Skin-Snip Microscopy, Polymerase Chain Reaction (PCR), ELISA test.

scholarly journals Tuberculosis in Birds: Insights into theMycobacterium aviumInfections

2011 ◽  
Vol 2011 ◽  
pp. 1-14 ◽  
Author(s):  
Kuldeep Dhama ◽  
Mahesh Mahendran ◽  
Ruchi Tiwari ◽  
Shambhu Dayal Singh ◽  
Deepak Kumar ◽  
...  
Keyword(s):  
Animal Health ◽  
Clinical Manifestations ◽  
Tuberculin Test ◽  
Rapid Identification ◽  
Molecular Techniques ◽  
Human Beings ◽  
Pet Birds ◽  
Polymerase Chain ◽  
World Organization

Tuberculosis, a List B disease of World Organization for Animal Health, caused byM. aviumorM. genavensepredominantly affects poultry and pet or captive birds. Clinical manifestations in birds include emaciation, depression and diarrhea along with marked atrophy of breast muscle. Unlike tuberculosis in animals and man, lesions in lungs are rare. Tubercular nodules can be seen in liver, spleen, intestine and bone marrow. Granulomatous lesion without calcification is a prominent feature. The disease is a rarity in organized poultry sector due to improved farm practices, but occurs in zoo aviaries. Molecular techniques like polymerase chain reaction combined with restriction fragment length polymorphism and gene probes aid in rapid identification and characterization of mycobacteria subspecies, and overcome disadvantages of conventional methods which are slow, labour intensive and may at times fail to produce precise results.M. avium subsp. aviumwith genotypeIS901+ andIS1245+ causes infections in animals and human beings too. The bacterium causes sensitivity in cattle to the tuberculin test. The paper discusses in brief theM. aviuminfection in birds, its importance in a zoonotic perspective, and outlines conventional and novel strategies for its diagnosis, prevention and eradication in domestic/pet birds and humans alike.

scholarly journals Q fever: A neglected disease of camels in Giza and Cairo Provinces, Egypt

2019 ◽  
Vol 12 (12) ◽  
pp. 1945-1950 ◽  
Author(s):  
Hend H. A. M. Abdullah ◽  
Hany A. Hussein ◽  
Khaled A. Abd El-Razik ◽  
Ashraf M. A. Barakat ◽  
Yousef A. Soliman
Keyword(s):  
Q Fever ◽  
Quantitative Polymerase Chain Reaction ◽  
Epidemiological Data ◽  
Acute Stage ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Molecular Tools ◽  
Blood Samples ◽  
Sensitive Tool ◽  
Polymerase Chain

Background and Aim: Q fever is a zoonotic disease caused by Coxiella burnetii. Cattle, sheep, and goat are the main reservoir of C. burnetii. In Egypt, the epidemiological data about C. burnetii in camels are limited. Therefore, the current study was conducted to identify C. burnetii infection in camels by different molecular tools and to estimate its seropositivity through the detection of anti-C. burnetii antibodies in camel sera. Materials and Methods: Blood samples were collected 112 from camels in Giza and Cairo Provinces, Egypt. All blood samples were screened by trans-quantitative polymerase chain reaction (trans-qPCR) for C. burnetii and positive samples subjected to standard PCR using the superoxide dismutase enzyme coding gene of C. burnetii. Sera of studied camels were examined for the presence of antibodies against C. burnetii using enzyme-linked immunosorbent assay. Results: Out of 112 camels, 19 were positive for C. burnetii by qPCR with an overall prevalence of 16.9% (18.6% in Giza and 15.1% in Cairo Provinces, respectively). The seroprevalence of anti-C. burnetii IgG antibodies in the examined camels was 4.5% (5/112). Conclusion: Trans-qPCR assay is a rapid and sensitive tool for the detection of C. burnetii in acute stage. Camels should be considered one of the major reservoirs for C. burnetii in Egypt.

scholarly journals Molecular Characterization of Salmonella Species Isolates from Some Hospitals in Jos, Nigeria

2021 ◽  
Vol 2 (2) ◽  
pp. 1-5
Author(s):  
S. B. Uneze ◽  
P. F. Chollom ◽  
Y. A. Agabi ◽  
D. J. Mawak ◽  
O. J. Egbere ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Characterization ◽  
Molecular Size ◽  
Molecular Techniques ◽  
The Other ◽  
Chain Reaction ◽  
Inva Gene ◽  
Polymerase Chain ◽  
Similarities And Differences

The conventional methods of identification of Salmonella involving microbiological enrichment and successive identification mostly are tedious, time consuming and not specific. Therefore, the aim of this study was to utilize molecular techniques to characterize Salmonella species isolates from some Hospitals in Jos, Nigeria. The 10 isolates collected from some Hospitals in Jos, Nigeria were screened for Salmonella using conventional biochemical methods. The positive isolates were identified using polymerase chain reaction (PCR) for discernment of invasion A (invA) gene at explicit molecular size (284 bp) utilizing explicit primers (forward and reverse). Sequencing of the invA gene was performed and the similarities and differences between our invA gene and published sequences on GenBank were assessed. Seven out of ten confirmed Salmonella species isolates were positive to the invA gene while the remaining three were negative. The homology level of nucleotide sequence (97.746%) demonstrated high similitude between the local isolates and the other sequences on GenBank. Molecular characterization of the Salmonella isolates provides data about the virulence of the pathogen just as its relatedness to different organisms which offer data about the genome of the organisms and are helpful for epidemiological examinations. Therefore, Molecular methods which enable the detection of virulent genes are extremely important surveillance tools that are required to assist in curbing the escalation of infections caused by Salmonella.

scholarly journals Molecular detection and phylogenetic tree of infectious laryngotracheitis virus in layers in Al-Diwaniyah province, Iraq

2019 ◽  
Vol 12 (4) ◽  
pp. 605-608 ◽  
Author(s):  
Furkan Alaraji ◽  
Hasan Hammadi ◽  
Alaa Abdulaziz Abed ◽  
Yahia Ismail Khudhair
Keyword(s):  
Dna Sequencing ◽  
Molecular Techniques ◽  
Infectious Laryngotracheitis Virus ◽  
P32 Gene ◽  
Field Samples ◽  
Infectious Laryngotracheitis ◽  
Polymerase Chain ◽  
Laryngotracheitis Virus ◽  
First Time

Background and Aim: Infectious laryngotracheitis (ILT) of chickens is a substantial issue to be studied in Iraq because this disease is one of the most highly contagious respiratory diseases in the world caused by a herpesvirus. However, in Iraq, the ILT virus (ILTV) infection and disease have yet not been confirmed in layers, so farm owners do not vaccinate these layers. The current study aimed to document the detection and characterization of ILTV in layer hens from Al-Diwaniyah city, for the first time in Iraq, using molecular techniques like polymerase chain reaction (PCR) and sequencing. Materials and Methods: Four layer farms (15,000 unvaccinated layers/farm) in Al-Diwaniyah province, Iraq, suffered a severe ILT outbreak, was diagnosed and reported by clinical and PCR tests. This disease has been reported in Iraq, and more recently, it began to show outbreaks in Al-Diwaniyah city. The current work opted to investigate the ILTV using PCR and DNA sequencing techniques. The study targeted the p32 gene of ILTV using pooled tracheal swabs and organs including the trachea, lung, and kidneys which were collected from dead and clinically infected chickens. Results: The analyses revealed that four of six suspected field samples showed positive results by PCR. The DNA sequencing results showed the homology of the amplified fragments with the studied gene. Conclusion: This study confirmed the presence of ILTV in hens with respiratory signs during the outbreak.

scholarly journals Low Frequency of Asymptomatic and Submicroscopic Plasmodial Infections in Urabá Region in Colombia

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Carolina Rodríguez Vásquez ◽  
Sebastián Barrera Escobar ◽  
Alberto Tobón-Castaño
Keyword(s):  
Low Frequency ◽  
Epidemiological Data ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Convenience Sample ◽  
Endemic Malaria ◽  
Blood Smears ◽  
Finger Prick ◽  
Polymerase Chain ◽  
Thick Smear

Background. A screening for malaria parasites was conducted with asymptomatic residents in Colombia.Methods. A descriptive study was carried out in December 2012 in four municipalities of Urabá region in Colombia. A convenience sample of 400 subjects was selected. Participants responded to a survey regarding epidemiological data and blood samples were taken from capillary blood obtained by finger prick for thick smear, rapid diagnostic test (RDT), and polymerase chain reaction (PCR).Results. 399 subjects aged 0.2-98 years were studied (median 22; 221 female (55%)). Episodes of malaria in the last year confirmed by thick film were reported by 47 participants (12%). In 399 samples tested by RDT 4 (1%) were positive (1 withP. falciparum, 3 withP. vivax), and 3 were confirmed by PCR. In 399 thick blood smears examined 5 (1.3%) were positive (2 withP. falciparum, 3 withP. vivax), and 3 were confirmed by PCR. In 227 samples, PCR showed 6 (2.6%) positive samples. The parasitaemia was below 1,440 parasites/μL. The best agreement between diagnoses was found between the RDT and thick blood smears (Kappa = 0.75).Conclusion. Plasmodial afebrile infection was found in 2% of the studied population, by three diagnostic methods, in residents from a low endemic malaria region in Colombia.

scholarly journals Semi-domesticated dogs as a potential reservoir for zoonotic hookworms in Bangkok, Thailand

2020 ◽  
Vol 13 (5) ◽  
pp. 909-915
Author(s):  
Jutamas Wongwigkan ◽  
Tawin Inpankaew
Keyword(s):  
Molecular Techniques ◽  
Parasitic Nematodes ◽  
Potential Source ◽  
Animal Populations ◽  
Potential Reservoir ◽  
Polymerase Chain ◽  
Temple Community ◽  
Hookworm Infections ◽  
Mitochondrial Cytochrome Oxidase

Background and Aim: Hookworms are parasitic nematodes that live in the small intestine of their mammalian hosts including humans, dogs, and cats. This study was conducted to determine the prevalence and perform genetic characterization of hookworms using molecular techniques and to elucidate the risk factors associated with hookworm infections among semi-domesticated dogs residing in temples in the Bangkok Metropolitan Area, Thailand. Materials and Methods: A total of 500 fecal samples were collected from semi-domesticated dogs from 91 temples in 48 districts of Bangkok. DNA was extracted and screened using internal transcribed spacer polymerase chain reaction-restriction fragment length polymorphism. In addition, samples positive for Ancylostoma ceylanicum were further characterized at the haplotype level based on the analysis of the mitochondrial cytochrome oxidase-1 gene (cox1). Results: The prevalence of hookworm infections in semi-domesticated dogs was 6.2% (31/500). Hookworm infections were detected in temple-community dogs in 12 of 48 districts (25.0%), with Bang Khen and Lak Si districts having the highest proportion of infected dogs (22.6%). Regarding molecular characterization of hookworm species, 21 positive samples (67.74%) were infected with A. ceylanicum and 10 (32.26%) with Ancylostoma caninum. Characterization of cox1 in A. ceylanicum isolates revealed the presence of a mixture of human and dog isolates. Conclusion: Semi-domesticated dogs act as a potential source of hookworm infections for human and animal populations in Bangkok, Thailand.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

1992 ◽  
Vol 67 (01) ◽  
pp. 063-065 ◽  
Author(s):  
Sherryl A M Taylor ◽  
Jacalyn Duffin ◽  
Cherie Cameron ◽  
Jerome Teitel ◽  
Bernadette Garvey ◽  
...  
Keyword(s):  
Nucleotide Substitution ◽  
Novel Mutation ◽  
Factor Ix ◽  
Causative Mutation ◽  
Coding Region ◽  
Body Of Knowledge ◽  
Polymerase Chain ◽  
Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

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