Detection of malaria with light microscopy and Nested polymerase chain reaction (Nested PCR) methods in peripheral blood expansions and investigation of the genetic diversity of Plasmodium species by 18S rRNA gene in Southeast of Iran

2019 ◽  
Vol 137 ◽  
pp. 103782
Author(s):  
Ahmad Taghdiri ◽  
Pooya Ghasemi Nejad Almani ◽  
Iraj Sharifi ◽  
Mohammad Ali Mohammadi ◽  
Samira Salari
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Light Microscopy ◽  
18S Rrna ◽  
Plasmodium Species ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals AMPLIFIKASI FRAGMEN GEN 18S rRNA PADA DNA METAGENOMIK MADU DENGAN TEKNIK PCR (POLYMERASE CHAIN REACTION)

2017 ◽  
Vol 7 ◽  
pp. 1
Author(s):  
Satriya Putra Prakoso ◽  
I Nengah Wirajana ◽  
I Wayan Suarsa
Keyword(s):  
Polymerase Chain Reaction ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Honey Sample ◽  
Rrna Gene ◽  
In Vitro Test ◽  
Chain Reaction ◽  
Metagenomic Dna ◽  
Polymerase Chain

The aim of this research was to amplificate 18S rRNA gene fragment from honey’s metagenomic DNA using Polymerase Chain Reaction (PCR). The honey sample was collected from Seraya Tengah village, Karangasem regency. The best result of primer design from in silico test was continued to in vitro test using PCR method. The optimum conditions for amplification was obtained as follows: pre-denaturation at 95oC for 3 minutes and continued with 30 of amplification cycle (denaturation at 95°C for 1 minutes, annealing at 55°C for 1 minutes and elongation at 72°C for 1 minutes) and the last step continued with extension process at 72°C for 2 minutes. The size of DNA fragment band of amplified product was about 100 bp which obtained from the honey’s metagenomic DNA.

scholarly journals Fatal Hymenolepis nana cestodiasis in a ring-tailed lemur (Lemur catta)

2021 ◽  
pp. 030098582110425
Author(s):  
Esther E. V. Crouch ◽  
Charlotte Hollinger ◽  
Stephanie Zec ◽  
Denise McAloose
Keyword(s):  
Polymerase Chain Reaction ◽  
18S Rrna ◽  
Lemur Catta ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Hymenolepis Nana ◽  
Chain Reaction ◽  
Fecal Shedding ◽  
Polymerase Chain ◽  
Common Parasite

The cestode Hymenolepis nana is a common parasite of humans and mice. Fecal shedding in the absence of clinical disease has previously been reported in ring-tailed lemurs ( Lemur catta). This report describes fatal, disseminated H. nana cestodiasis infection in an aged ring-tailed lemur in a zoological collection. The parasites were associated with severe multifocal to coalescing and regionally extensive pyogranulomatous hepatitis and moderate multifocal pneumonia. The morphology of the parasites was highly unusual. Profiles were variably sized, ellipsoid to irregularly serpiginous, lined by a thin tegument, and filled with lightly eosinophilic fibrillar stroma and numerous, round basophilic cells. Polymerase chain reaction targeting a portion of the 18S rRNA gene and DNA sequencing of the amplicon showed 100% homology with H. nana.

scholarly journals How prevalent is Plasmodium malariae in Rondônia, Western Brazilian Amazon?

2000 ◽  
Vol 33 (5) ◽  
pp. 489-492 ◽  
Author(s):  
Marisa Torres Vidal Cavasini ◽  
Weber Luidi Ribeiro ◽  
Fumihiko Kawamoto ◽  
Marcelo Urbano Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Species ◽  
Brazilian Amazon ◽  
Plasmodium Malariae ◽  
Mixed Species ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Potential Impact ◽  
And Control

We have compared results of Plasmodium species identification obtained with conventional on-site microscopy of Giemsa-stained thick smears (GTS) and a semi-nested polymerase chain reaction (PCR) in 96 malaria patients from Rondônia, Western Brazilian Amazon. Mixed-species infections were detected by PCR in 30% patients, but no such case had been found on GTS. Moreover, P. malariae infections were detected in 9 of 96 patients (10%) by PCR, but were not identified by local microscopists. The potential impact of species misidentification on malaria treatment and control is discussed.

scholarly journals Detection of fish allergen by droplet digital PCR

2019 ◽  
Vol 7 (4) ◽  
Author(s):  
Cinzia Daga ◽  
Simona Cau ◽  
Maria Giovanna Tilocca ◽  
Barbara Soro ◽  
Aldo Marongiu ◽  
...  
Keyword(s):  
18S Rrna ◽  
Annealing Temperature ◽  
Pcr Primers ◽  
Digital Pcr ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Chain Reaction ◽  
European Regulation ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Fish is one of fourteen allergens that must be highlighted on the label within the ingredients list. It should be noted that the European regulation, is very restrictive to allergens with zero tolerance. Therefore it is important to establish sensitive and specific methods for detecting fish allergen. Applicability to detect and quantify fish allergen by droplet digital polymerase chain reaction (ddPCR) has been evaluated in this work. Genomic DNA of three fish species belonging to the most common fish families were analyzed. PCR primers were designed to amplify a 166 bp region of the 18S rRNA gene. Comparative studies were performed to establish the optimal primer and probe concentrations.  Annealing temperature was determined by using thermal gradient. The results have shown good applicability of the optimized 18S rRNA gene-method to detect and quantify small amounts of the target in all samples analyzed. However, validation studies are needed in order to apply ddPCR technology for routine allergens analysis.  

scholarly journals Babesia vogeli in dogs from Rio Branco, South-west Amazonia, Brazil

2021 ◽  
Vol 42 (6) ◽  
pp. 3527-3534
Author(s):  
Mayara Marques Pereira Fernandes ◽  
◽  
Marcelo Renan Serrate Rodrigues ◽  
Jessica Damiana Marinho Valente ◽  
Marcelli Pascoal Nogueira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Detection ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Babesia Vogeli ◽  
Physical Examinations ◽  
Northern Brazil ◽  
Polymerase Chain

This is the first report of Babesia vogeli molecular detection in dogs from the state of Acre, northern Brazil. This study aimed to perform the molecular detection of Babesia vogeli in dogs in the municipality of Rio Branco, Acre. Blood samples were collected from 47 dogs presenting with clinical signs comparable to hemoparasitosis. These were dogs which were attended in veterinary clinics from Rio Branco municipality, Acre. Physical examinations, packed cell volume (PCV) determination, platelet number estimation, hemoparasite investigation in the blood (collected from the pinna and peripheral blood), and polymerase chain reaction (PCR) for piroplasm based on the 18S rRNA gene, were performed. One dog (1/47, 2.1%; CI 95%: 0.1-11.3%) tested positive to Babesia vogeli in the polymerase chain reaction (PCR) assay for piroplasms and the resulting sequence showed 100% identity with Babesia vogeli isolates deposited in GenBank®. Co-infection with Ehrlichia spp. was also observed by direct examination (via blood smear). The clinical and hematological alterations observed in the positive animal were anorexia, dehydration, white mucous membranes, anemia and thrombocytopenia.

scholarly journals Designing primers potentially specific to Entamoeba gingivalis genes

2015 ◽  
Vol 28 (3) ◽  
pp. 159-163
Author(s):  
Jagoda Abramek
Keyword(s):  
Polymerase Chain Reaction ◽  
Oral Cavity ◽  
Light Microscopy ◽  
Genital Tract ◽  
18S Rrna ◽  
Acute Osteomyelitis ◽  
Chain Reaction ◽  
Human Oral Cavity ◽  
Pulmonary Abscess ◽  
Polymerase Chain

Abstract Entamoeba gingivalis normally exists in the human oral cavity, namely in the gums, and brings about some specific diseases. However, it can also trigger some more serious illnesses. Among these are infections of the genital tract, acute osteomyelitis of the mandible and pulmonary abscess. Entamoeba gingivalis identification by light microscopy is difficult, hence polymerase chain reaction (PCR) is used. The contemporary primers for PCR are complement to 18S rRNA. This article informs the reader of the process that was involved in designing new primers for three genes which were thought to be present on the Entamoeba gingivalis genome, but their sequences were unknown. The newly obtained sequences of primers have better properties for identification purposes, compared to these which are currently used.

An Oligonucleotide-Ligation Assay for the Differentiation between Cyclospora and Eimeria spp. Polymerase Chain Reaction Amplification Products

1999 ◽  
Vol 62 (6) ◽  
pp. 682-685 ◽  
Author(s):  
KAREN C. JINNEMAN ◽  
JUNE H. WETHERINGTON ◽  
WALTER E. HILL ◽  
CURTIS J. OMIESCINSKI ◽  
ANN M. ADAMS ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Amplification ◽  
Pcr Amplification ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Eimeria Spp ◽  
Pcr Rflp ◽  
Oligonucleotide Ligation Assay ◽  
Polymerase Chain

An oligonucleotide-ligation assay (OLA) was developed and compared to a restriction fragment length polymorphism (RFLP) test for distinguishing between 294-bp polymerase chain reaction (PCR) amplification products of the 18S rRNA gene from Cyclospora and Eimeria spp. The PCR/OLA correctly distinguished between three Cyclospora, three E. tenella, and one E. mitis strains and the ratio of positive to negative spectrophotometric absorbance (A490) values for each strain ranged from 4.086 to 15.280 (median 9.5). PCR/OLA provides a rapid, reliable, spectrophotometric alternative to PCR/RFLP.

A 23S rDNA-Targeted Polymerase Chain Reaction–Based System for Detection of Staphylococcus aureus in Meat Starter Cultures and Dairy Products

1999 ◽  
Vol 62 (10) ◽  
pp. 1150-1156 ◽  
Author(s):  
JOACHIM A. STRAUB ◽  
CHRISTIAN HERTEL ◽  
WALTER P. HAMMES
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Skim Milk ◽  
Starter Cultures ◽  
23S Rrna ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Selective Enrichment ◽  
Polymerase Chain

A polymerase chain reaction–based system for detection of Staphylococcus aureus was developed. The system consisted of the following components: (i) selective enrichment, (ii) DNA isolation, (iii) amplification of DNA with primers targeted against the 23S rRNA gene, and (iv) evaluation of the specificity of the polymerase chain reaction by Southern hybridization and nested polymerase chain reaction. The method achieved a high degree of sensitivity and unambiguity as required for the detection of contaminants in food starter preparations. The method permitted detection of Staphylococcus aureus in preparations of meat starter cultures containing Staphylococcus carnosus either alone or in combination with lactobacilli, pediococci, and/or Kocuria varians. Detection limits were sufficiently low to show within 12 h the presence of 100 CFU of S. aureus in starter preparations containing 1010 CFU of S. carnosus. The system was also applied to dried skim milk and cream. For detection without selective enrichment, a protocol was developed and permitted detection of 120 CFU of S. aureus in 1 ml of cream within 6 h. With nested polymerase chain reaction, the detection limit was decreased by one order of magnitude.

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