scholarly journals Detection of the jaundice-related G71R mutation in the UGT1A1 gene by denaturing high performance liquid chromatography (DHPLC)

2009 ◽  
Vol 49 (1) ◽  
pp. 1
Author(s):  
Retno Sutomo ◽  
Sunartini Hapsara ◽  
Suryono Yudha Patria ◽  
Hajime Nakamura
Keyword(s):  
Sequence Analysis ◽  
Gold Standard ◽  
High Performance ◽  
Population Based ◽  
Sequencing Analysis ◽  
Chain Reaction ◽  
Unconjugated Hyperbilirubinemia ◽  
Ugt1a1 Gene ◽  
Exon 1 ◽  
Dhplc Analysis

Background  The  G71R mutation in the UGT1A1 gene has  beenassociated with neonatal jaundice  and  other  cases  of  hereditary,unconjugated hyperbilirubinemia in several Asian populations.Currently,  DNA  sequencing  is  the  only  method  available  toidentify the mutation, which can be time- and  labor-intensive,particularly for such projects  as  population-based genetic studies.A relatively new method, denaturing high performance liquidchromatography (DHPLC),  is  increasingly used to  detect  variousmutations.Objective  The  aim  of  the present study was to investigate theability of DHPLC to  detect  the G71R mutation, in comparisonwith the gold standard of sequencing analysis.Methods Seventy-two infants were enrolled. Following genomicDNA  extraction, exon 1 of the UGT1A1 gene was amplified  bypolymerase chain reaction (PCR). Afterwards, the G71R mutationwas simultaneously,  and  blindly, determined in all subjects  byDHPLC and sequence analysis.  The  performance  of  the DHPLCanalysis, compared  to  the sequence analysis, was assessed in termsof  sensitivity  and  specificity.Results DHPLC detected the G71 R mutation in  31  individuals.Of  these,  26  were heterozygous and 5 were homozygous for themutation. This method did not find the mutation in  41  otherindividuals. Sequence analysis produced identical results for allindividuals.Conclusion DHPLC analysis  is  capable  of  detecting the G71Rmutation  in  the  UGT1A1  with  a degree  of  sensitivity  andspecificity  (100%  each)  that  is  comparable to sequencing analysis.

scholarly journals UGT1A1 gene polymorphisms and jaundice in Indonesian neonates

2019 ◽  
Vol 59 (3) ◽  
pp. 150-6 ◽  
Author(s):  
Rinawati Rohsiswatmo ◽  
Radhian Amandito ◽  
Andiani Wanda Putri ◽  
Nilam Sartika ◽  
Amarila Malik
Keyword(s):  
Neonatal Intensive Care ◽  
Asian Population ◽  
Restriction Enzymes ◽  
Total Serum ◽  
Unconjugated Hyperbilirubinemia ◽  
Ugt1a1 Gene ◽  
Racial Variations ◽  
Exon 1 ◽  
Polymerase Chain ◽  
Heterozygous Form

Background Uridine diphospho-glucuronocyltransferase 1A1 (UGT1A1) polymorphisms are a risk factor for unconjugated hyperbilirubinemia in neonates. UGT1A1 polymorphisms decrease bilirubin conjugation, thus causing hyperbilirubinemia. A variety of polymorphisms have been reported, with UGT1A1*60 and UGT1A1*6 especially prominent in the Asian population. Hyperbilirubinemia polymorphism studies are lacking in Indonesian populations. Objective To identify UGT1A1*60 and UGT1A1*6 profiles in Indonesian populations of heterogeneous ethnicity. Methods We enrolled 42 jaundiced neonates who were born from January to April 2017 and treated in the Neonatal Intensive Care Unit of our national referral center, Cipto Mangunkusumo Hospital, Jakarta, Indonesia. Genetic mutations *60 of exon 1 and *6 of the promoter region were analyzed by polymerase chain reaction – restriction fragment length polymorphism methods, with DraI and AvaII as restriction enzymes, respectively. Clinical data including total serum bilirubin and racial information were obtained by medical records and interviews with parents. Results There were no homozygous mutations of UGT1A1*6, but 4.8% of subjects were heterozygous. As for UGT1A1*60, 4.8% were heterozygous and 95.2% were homozygous. Racial variations were not observed for UGT1A1*60, while Betawi descendents were found to have many heteroygous forms of UGT1A1*6. Conclusion Polymorphisms of the UGT1A1 gene were found in Indonesian neonates. Some ethnicities also showed increased tendency towards its incidence, such as the heterozygous form of  UGT1A1*6.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

scholarly journals Evaluation of four different HPLC devices for hemoglobinopathy screening

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Müjgan Ercan Karadağ ◽  
Emiş Deniz Akbulut ◽  
Esin Avcı ◽  
Esra Fırat Oğuz ◽  
Saadet Kader ◽  
...  
Keyword(s):  
Gold Standard ◽  
High Performance ◽  
Public Health Problem ◽  
Reference Value ◽  
Comparison Study ◽  
Gold Standard Method ◽  
Blood Samples ◽  
Systematic Biases ◽  
Hemoglobin Variants ◽  
Thalassemia Screening

AbstractObjectiveHemoglobinopathies are a common public health problem in Turkey. In the screening of these disorders in population, cation-exchange high performance liquid chromatography (HPLC) is accepted as the gold standard method. In this study, the aim was to assess four different HPLC devices used in hemoglobinopathy screening.Materials and methodsA total of 58 blood samples were analyzed with four different HPLC methods (Bio-Rad variant II, Agilent 1100, Tosoh G8 and Trinity Ultra2 trademarks).ResultsThe comparison study demonstrated a good correlation between the results of each HPLC analyzer and the reference value obtained by averaging all the HbA2 results belonging to the methods tested in the study [ (Tosoh G8 (r=0.988), Bio-Rad variant II (r=0.993), Agilent 1100 (r=0.98) and Trinity Ultra2 (r=0.992) ]. HbA2 determination in the presence of HbE was interfered in both Bio-Rad variant II and Tosoh G8.ConclusionThe analyzers were found to have compatible HbA2 results but with accompanying different degrees of proportional and systematic biases. HPLC analyzers may be affected by different hemoglobin variants at different HbA2 concentrations, which is an important point to take into consideration during the evaluation of HbA2 results in thalassemia screening.

scholarly journals Performance of Seven Consumer Sleep-Tracking Devices Compared with Polysomnography

SLEEP ◽  
2020 ◽  
Author(s):  
Evan D Chinoy ◽  
Joseph A Cuellar ◽  
Kirbie E Huwa ◽  
Jason T Jameson ◽  
Catherine H Watson ◽  
...  
Keyword(s):  
Performance Measures ◽  
Gold Standard ◽  
High Performance ◽  
Sleep Stage ◽  
Sleep Laboratory ◽  
Sleep Assessment ◽  
Sleep Condition ◽  
Different Populations ◽  
Tracking Devices ◽  
Better Than

Abstract Study Objectives Consumer sleep-tracking devices are widely used and becoming more technologically advanced, creating strong interest from researchers and clinicians for their possible use as alternatives to standard actigraphy. We therefore tested the performance of many of the latest consumer sleep-tracking devices, alongside actigraphy, versus the gold-standard sleep assessment technique, polysomnography (PSG). Methods In total, 34 healthy young adults (22 women; 28.1 ± 3.9 years, mean ± SD) were tested on three consecutive nights (including a disrupted sleep condition) in a sleep laboratory with PSG, along with actigraphy (Philips Respironics Actiwatch 2) and a subset of consumer sleep-tracking devices. Altogether, four wearable (Fatigue Science Readiband, Fitbit Alta HR, Garmin Fenix 5S, Garmin Vivosmart 3) and three non-wearable (EarlySense Live, ResMed S+, SleepScore Max) devices were tested. Sleep/wake summary and epoch-by-epoch agreement measures were compared with PSG. Results Most devices (Fatigue Science Readiband, Fitbit Alta HR, EarlySense Live, ResMed S+, SleepScore Max) performed as well as or better than actigraphy on sleep/wake performance measures, while the Garmin devices performed worse. Overall, epoch-by-epoch sensitivity was high (all ≥0.93), specificity was low-to-medium (0.18-0.54), sleep stage comparisons were mixed, and devices tended to perform worse on nights with poorer/disrupted sleep. Conclusions Consumer sleep-tracking devices exhibited high performance in detecting sleep, and most performed equivalent to (or better than) actigraphy in detecting wake. Device sleep stage assessments were inconsistent. Findings indicate that many newer sleep-tracking devices demonstrate promising performance for tracking sleep and wake. Devices should be tested in different populations and settings to further examine their wider validity and utility.

Frequency of the delta Phe508 mutation and correlation with XV.2c/KM-19 haplotypes in an American population of cystic fibrosis patients: results of a collaborative study

1990 ◽  
Vol 36 (10) ◽  
pp. 1741-1746 ◽  
Author(s):  
W E Highsmith ◽  
G L Chong ◽  
H T Orr ◽  
T R Perry ◽  
D Schald ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Haplotype Analysis ◽  
Screening Program ◽  
Collaborative Study ◽  
Population Based ◽  
Carrier Status ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Phenylalanine Residue ◽  
Test Result

Abstract The cystic fibrosis (CF) gene has been recently cloned, and a deletion of 3 basepairs (bp) of DNA was found on most of the CF chromosomes. This deletion leads to the synthesis of a protein that lacks a phenylalanine residue at position 508. Using two polymerase chain reaction protocols to study the frequency of this mutation in a series of 192 CF patients, we found the mutation on 72% of affected chromosomes. We then used this value to calculate the predictive value of a negative test result in a population-based screening program for CF carrier status. Haplotype analysis with the polymorphic markers XV.2c and KM-19 on 239 CF chromosomes revealed that 90.7% of CF chromosomes with the deletion had a single haplotype. This haplotype was also associated with 60.4% of CF chromosomes with unknown mutations. These values can be used to calculate the probability of whether an individual from the general population is a carrier of any CF mutation.

scholarly journals Plasma Lutein, a Nutritional Biomarker for Development of Advanced Age-Related Macular Degeneration: The Alienor Study

Nutrients ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 2047
Author(s):  
Bénédicte M. J. Merle ◽  
Audrey Cougnard-Grégoire ◽  
Jean-François Korobelnik ◽  
Wolfgang Schalch ◽  
Stéphane Etheve ◽  
...  
Keyword(s):  
Macular Degeneration ◽  
High Performance ◽  
Population Based ◽  
Age Related Macular Degeneration ◽  
Cox Proportional Hazard ◽  
Age Related ◽  
Cox Proportional Hazard Models ◽  
Adjusted Model ◽  
Reduced Risk

Lutein and zeaxanthin may lower the risk of age-related macular degeneration (AMD). We evaluated the associations of plasma lutein and zeaxanthin with the incidence of advanced AMD in the Alienor study (Antioxydants Lipides Essentiels Nutrition et Maladies Oculaires). Alienor study is a prospective population-based cohort of 963 residents of Bordeaux, France, who were 73 years or older at baseline (2006–2008). The present study included 609 participants with complete ophthalmologic and plasma carotenoids data. Examinations were performed every two years over an eight-year period (2006 to 2017). Plasma lutein and zeaxanthin were determined at baseline from fasting blood samples using high-performance liquid chromatography. Cox proportional hazard models were used to assess associations between plasma lutein, zeaxanthin, and their (total cholesterol (TC) + triglycerides (TG)) ratios with AMD. Among the 609 included participants, 54 developed advanced incident AMD during a median follow-up time of 7.6 years (range 0.7 to 10.4). Participants with higher plasma lutein had a reduced risk for incident advanced AMD in the fully adjusted model (HR = 0.63 per 1-SD increase (95% CI, 0.41–0.97), p = 0.03). A similar association was observed using the lutein/(TC + TG) ratio (HR = 0.59 (95% CI, 0.39–0.90), p = 0.01). No associations were evidenced for other carotenoids. Higher plasma lutein was associated with a 37% reduced risk of incident advanced AMD.

scholarly journals The Identification of Mutation in BMP15 Gene Associated with Litter Size in Xinjiang Cele Black Sheep

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 668
Author(s):  
Zhi-gang Niu ◽  
Jin Qin ◽  
Yao Jiang ◽  
Xiang-Dong Ding ◽  
Yu-gong Ding ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Litter Size ◽  
Base Change ◽  
Wild Type ◽  
Wild Genotype ◽  
Exon 2 ◽  
Black Sheep ◽  
Morphogenetic Protein ◽  
Gene Sequence Analysis ◽  
Exon 1

The Bone Morphogenetic Protein 15 (BMP15) gene is known to have multiple single-nucleotide polymorphism sites associated with sheep fecundity. This study used gene sequence analysis and mutation detection assays for BMP15 by using 205 blood samples of ewes with known lambing records. Sequence analysis showed that mutation B1 missed the CTT base in exon 1 at positions 28–30, leading to a leucine deletion in the BMP15 protein. Litter size of ewes differed significantly between BB and B+ genotypes of B1 (p < 0.05); however, the differences between wild genotype (++) and homozygous (BB) or wild genotype (++) and heterozygous (B+) were not significant (p > 0.05). Another mutation, T755C, is a T-to-C base change at position 755 of exon 2, resulting in leucine replacement by proline at this position of the BMP15 protein (p.L252P). Two genotypes were identified in the flock: heterozygous (E+) and wild-type genotype (++). Ewes with heterozygous (E+) p.L252P had significantly larger litter sizes than those with the wild-type genotype (p < 0.05). Comprehensive analysis suggests that p.L252P is a mutation that affects fecundity in Cele black sheep.

scholarly journals Utility of forensic detection of rabies virus in decomposed exhumed dog carcasses

2015 ◽  
Vol 86 (1) ◽  
Author(s):  
Wanda Markotter ◽  
Jessica Coertse ◽  
Kevin Le Roux ◽  
Joey Peens ◽  
Jacqueline Weyer ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Gold Standard ◽  
Diagnostic Method ◽  
Diagnostic Testing ◽  
Domestic Dogs ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Brain Material ◽  
Polymerase Chain ◽  
Genomic Material

This report describes four suspected rabies cases in domestic dogs that were involved inhuman exposures. In all these cases, the animals were buried for substantial times beforerabies testing was performed. Animal rabies is endemic in South Africa and domestic dogsare the main vector for transmission to humans. Diagnosis of rabies in humans is complicated,and diagnosis in the animal vector can provide circumstantial evidence to support clinicaldiagnosis of rabies in humans. The gold standard diagnostic method, fluorescent antibodytest (FAT), only delivers reliable results when performed on fresh brain material and thereforedecomposed samples are rarely submitted for diagnostic testing. Severely decomposed brainmaterial was tested for the presence of rabies virus genomic material using a quantitativereal-time reverse transcription polymerase chain reaction (q-real-time RT-PCR) assaywhen conventional molecular methods were unsuccessful. This may be a useful tool in theinvestigation of cases where the opportunity to sample the suspected animals post mortem wasforfeited and which would not be possible with conventional testing methodologies becauseof the decomposition of the material.

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