scholarly journals Molecular identification and genotyping of Acinetobacter baumannii isolated from burn patients by PCR and ERIC-PCR

2019 ◽  
Vol 5 ◽  
pp. 205951311983136 ◽  
Author(s):  
Faezeh Falah ◽  
Leili Shokoohizadeh ◽  
Maryam Adabi
Keyword(s):  
Acinetobacter Baumannii ◽  
Epidemiological Studies ◽  
Burn Patients ◽  
Chain Reaction ◽  
Burn Wound ◽  
Accurate Identification ◽  
Microbiological Test ◽  
Major Burn ◽  
Screening And Identification ◽  
Polymerase Chain

Background: Acinetobacter baumannii is one of the most important agents of hospital infections. Rapid and accurate identification and genotyping of A. baumannii is very important, especially in burn hospitals in order to prevent the spread of related nosocomial infections and to further epidemiological studies. Material and methods: For two months, 82 A. baumannii isolates were collected from burn wound swabs of patients in a major burn hospital in Tehran. A. baumannii isolates were identified by conventional microbiological test and polymerase chain reaction (PCR) using the primers of blaOXA-51 gene, while the genetic linkage of A. baumannii isolates was investigated by enterobacterial repetitive intragenic consensus (ERIC)-PCR technique. Similarity, a cut-off of ⩾ 95% was considered for classifying the genotypes. Results: The molecular test (PCR) confirmed 97.56% of phenotypic results for the detection of A. baumannii isolates. ERIC-PCR results revealed 14 different ERIC patterns (ERIC-types) including 11 common types and three unique types. Conclusion: Our findings show that we can simply and quickly detect A. baumannii isolates by PCR using blaOXA genes and genetic diversity by ERIC-PCR, respectively. These rapid and simple techniques for the routine screening and identification of clinical A. baumannii isolates could be useful with epidemic potential.

A New Resuscitation Formula Based on Burn Index Provides More Reliable Prediction for Fluid Requirement in Adult Major Burn Patients

2021 ◽  
Author(s):  
Jianglin Tan ◽  
Junyi Zhou ◽  
Ning Li ◽  
Lili Yuan ◽  
Gaoxing Luo
Keyword(s):  
Fluid Resuscitation ◽  
Linear Regression Analysis ◽  
Fluid Volume ◽  
Multivariate Linear Regression Analysis ◽  
Burn Patients ◽  
Full Thickness ◽  
Burn Wound ◽  
Major Burn ◽  
Fluid Requirement ◽  
Actual Volume

Abstract The Third Military Medical University (TMMU) formula is widely used in fluid resuscitation in China. However, the actual volume needs usually exceed the prediction provided by the TMMU formula in major burn patients with a high proportion of full-thickness burn wounds. This retrospective study included 149 adult major burn patients (≥40% TBSA) who were admitted to the Burn Department, Southwest Hospital from 2014 to 2020 and received appropriate fluid resuscitation by the TMMU protocol. The actual volume infused in the first 48 hours postburn was compared to the estimation by the TMMU formula. A new fluid volume prediction formula was developed by multivariate linear regression analysis. The mean fluid requirements were 2.35 ml/kg/% TBSA and 1.75 ml/kg/% TBSA in the first and second 24 hours postburn, respectively. The TMMU formula underestimated the fluid requirement, and its prediction accuracy was 54.1% and 25.8% for the first and second 24 hours, respectively. The proportion of full-thickness burn wound was found to be associated with the fluid requirements postburn. A revised multifactorial formula consisting of the burn index, body weight, and inhalation injury was developed. Using the revised formula, the prediction reliability of resuscitation fluid volume improved to 65.3% and 61.1% in the first and second 24 hours, respectively. The TMMU formula showed low accuracy in predicting fluid requirements among major burn patients. A revised formula based on burn index was developed to provide better guidance for initiative fluid resuscitation for major burns by the TMMU protocol.

Diagnostic Value of Multiplex Polymerase Chain Reaction in Detection of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from Sepsis in Pediatrics

2021 ◽  
Vol 15 ◽  
Author(s):  
Sara Galeb ◽  
Maysaa El Sayed Zaki ◽  
Raghdaa Shrief ◽  
Rasha Hassan ◽  
Mohamed Anies
Keyword(s):  
Polymerase Chain Reaction ◽  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Stenotrophomonas Maltophilia ◽  
Multiplex Polymerase Chain Reaction ◽  
Diagnostic Value ◽  
Blood Cultures ◽  
Chain Reaction ◽  
Polymerase Chain

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. Objectives: The present study aimed to evaluate the multiplex Polymerase Chain Reaction (PCR) to detect Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children, with hospital-acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR, with primers specific to the 3 tested bacteria. Results: Multiplex PCR was positive in 96% of isolates, and 4 isolates had negative results. False positive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa, and 27 isolates of Stenotrophomonas maltophilia. Compared to the biochemical identification, the diagnostic value of the multiplex PCR revealed 96.04% sensitivity, 96.9% specificity, 97.00%, positive predictive value, 96.00% negative predictive value, and 96.50% accuracy. Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific, and accurate. The accuracy differs according to the organisms, with 100% accuracy for Acinetobacter baumannii.

Determination of a T/G polymorphism at nucleotide 3206 of the apolipoprotein C III gene by amplification refractory mutation system

1994 ◽  
Vol 40 (12) ◽  
pp. 2235-2239 ◽  
Author(s):  
M Y Tsai ◽  
N Q Hanson ◽  
K R Copeland ◽  
I Beheshti ◽  
U Garg
Keyword(s):  
Epidemiological Studies ◽  
Amplification Refractory Mutation System ◽  
Chain Reaction ◽  
Apolipoprotein C ◽  
Significant Difference ◽  
Mutation System ◽  
Polymerase Chain ◽  
Exon 4 ◽  
And Control

Abstract We used the amplification refractory mutation system (ARMS)--a polymerase-chain-reaction-based method--to determine the 3206 T-to-G polymorphism on exon 4 of the apolipoprotein (apo) C III gene. Apo C III is an inhibitor of the enzyme lipoprotein lipase (EC 3.1.1.34). Previous studies have demonstrated that a polymorphism at nucleotide 3175 on exon 4 of this gene is associated with hypertriglyceridemia. We studied 45 hypertriglyceridemic and 46 age-matched controls for the 3206 T-to-G polymorphism. The results showed a significant difference in the distribution of the genotypes with respect to this allele between the hypertriglyceridemic and control individuals. We also determined the presence of the SacI site at nucleotide 3175 in these same individuals and found no significant difference in SacI genotypes between the two groups. This study reaffirms the usefulness of ARMS as a simple, reliable method for detecting mutations and polymorphisms in clinical and epidemiological studies.

scholarly journals DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL

2014 ◽  
Vol 56 (5) ◽  
pp. 391-395 ◽  
Author(s):  
Herintha Coeto Neitzke-Abreu ◽  
Kárin Rosi Reinhold-Castro ◽  
Mateus Sabaini Venazzi ◽  
Regiane Bertin de Lima Scodro ◽  
Alessandra de Cassia Dias ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Natural Infection ◽  
Epidemiological Studies ◽  
Leishmania Infection ◽  
Southern Brazil ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sensitivity Specificity

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

scholarly journals Optimization and Application of a Quantitative Polymerase Chain Reaction Assay to Detect Diaporthe Species in Soybean Plant Tissue

Plant Disease ◽  
2016 ◽  
Vol 100 (8) ◽  
pp. 1669-1676 ◽  
Author(s):  
Brian Kontz ◽  
Sajag Adhikari ◽  
Senthil Subramanian ◽  
Febina M. Mathew
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Stem Canker ◽  
Taqman Probe ◽  
Soybean Plant ◽  
Chain Reaction ◽  
Accurate Identification ◽  
Field Samples ◽  
Polymerase Chain ◽  
Soybean Plants

Diaporthe caulivora and D. longicolla are the causal agents of stem canker of soybean (Glycine max L.). Accurate identification of stem canker pathogens upon isolation from infected soybean plants is difficult and unreliable based on morphology. In this study, two TaqMan probe-based quantitative polymerase chain reaction (qPCR) assays were optimized for detection of D. caulivora and D. longicolla in soybean plants. The assays used previously reported D. caulivora-specific (DPC-3) and D. longicolla-specific (PL-3) probe/primer sets. The sensitivity limit of the two assays was determined to be over a range of 100 pg to 10 fg of pure D. caulivora and D. longicolla genomic DNA. The qPCR assays were validated with plant samples collected from commercial soybean fields. The PL-3 set detected D. longicolla in soybean plants collected from the fields (quantification cycle value <35), which was confirmed by isolation on potato dextrose agar (PDA). D. caulivora was detected only in low levels (quantification cycle value <40) by DPC-3 set in a few of the symptomatic field samples, although the pathogen was not isolated on PDA. The qPCR assays were also useful in quantitatively phenotyping soybean plants for resistance to D. caulivora and D. longicolla under greenhouse conditions.

Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing

2013 ◽  
Vol 89 (1) ◽  
pp. 118-123 ◽  
Author(s):  
L. Sadaow ◽  
C. Tantrawatpan ◽  
P.M. Intapan ◽  
V. Lulitanond ◽  
T. Boonmars ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Trichinella Spiralis ◽  
Large Subunit ◽  
Pcr Amplification ◽  
Epidemiological Studies ◽  
Chain Reaction ◽  
Target Region ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Zoonotic Parasites

AbstractNematodes of the genus Trichinella which infect wildlife and domestic animals show a cosmopolitan distribution. These zoonotic parasites are the aetiological agents of a severe human disease, trichinellosis. Twelve taxa are recognized in the Trichinella genus, but they cannot be identified by morphology since they are sibling species/genotypes. For epidemiological studies, it is extremely important to identify each taxon since they have different distribution areas and host ranges. In the present study, polymerase chain reaction (PCR) amplification of the mitochondrial large subunit ribosomal RNA (lsu-RNA) gene coupled with a pyrosequencing technique was developed to distinguish among four Trichinella species: Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis. A PCR method was used to amplify the lsu-RNA of Trichinella sp. larvae in mouse muscles and single larvae collected from infected muscles by digestion. The results show that the four Trichinella species can be distinguished by using 26 nucleotides in the target region and the method is sensitive enough to identify individual larvae. The pyrosequencing provides a simple, rapid and high-throughput tool for the differentiation of Trichinella species.

scholarly journals Mating Type Locus-Specific Polymerase Chain Reaction Markers for Differentiation of Pyrenophora teres f. teres and P. teres f. maculata, the Causal Agents of Barley Net Blotch

2010 ◽  
Vol 100 (12) ◽  
pp. 1298-1306 ◽  
Author(s):  
Shunwen Lu ◽  
Gregory J. Platz ◽  
Michael C. Edwards ◽  
Timothy L. Friesen
Keyword(s):  
Polymerase Chain Reaction ◽  
Mating Type ◽  
Pathogen Detection ◽  
Epidemiological Studies ◽  
Pyrenophora Teres ◽  
Nucleotide Polymorphisms ◽  
Net Blotch ◽  
Chain Reaction ◽  
Type Locus ◽  
Polymerase Chain

Fourteen single nucleotide polymorphisms (SNPs) were identified at the mating type (MAT) loci of Pyrenophora teres f. teres (Ptt), which causes net form (NF) net blotch, and P. teres f. maculata (Ptm), which causes spot form (SF) net blotch of barley. MAT-specific SNP primers were developed for polymerase chain reaction (PCR) and the two forms were differentiated by distinct PCR products: PttMAT1-1 (1,143 bp) and PttMAT1-2 (1,421 bp) for NF MAT1-1 and MAT1-2 isolates; PtmMAT1-1 (194 bp) and PtmMAT1-2 (939 bp) for SF MAT1-1 and MAT1-2 isolates, respectively. Specificity was validated using 37 NF and 17 SF isolates collected from different geographic regions. Both MAT1-1 and MAT1-2 SNP primers retained respective specificity when used in duplex PCR. No cross-reactions were observed with DNA from P. graminea, P. tritici-repentis, or other ascomycetes, or barley. Single or mixed infections of the two different forms were also differentiated. This study provides the first evidence that the limited SNPs at the MAT locus are sufficient for distinguishing closely related heterothallic ascomycetes at subspecies levels, thus allowing pathogenicity and mating type characteristics of the fungus to be determined simultaneously. Methods presented will facilitate pathogen detection, disease management, and epidemiological studies.

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