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1.
Children (Basel) ; 10(6)2023 Jun 01.
Article En | MEDLINE | ID: mdl-37371229

(1) Background: Fetal growth restriction (FGR) increases the risk of adverse neurodevelopmental outcomes, especially in preterm newborns. This study aims to describe the behavioral results of FGR at 6 years of age and to demonstrate the relationship of certain predictive factors with this development. (2) Methods: This retrospective cohort study included 70 children born in 2015 at the University Hospital Carlos Haya, Málaga, Spain who had been exposed to FGR during pregnancy; neonatal and infant data were recorded retrospectively. Children were assessed prospectively at 6 years of age by means of a strengths and difficulties questionnaire (SDQ) to study behavioral outcomes. (3) Results: We demonstrated that there are higher behavioral disability rates in children exposed to FGR during pregnancy and, in particular, high rates of hyperactivity or conduct problems. We also proved a negative relationship between the birth weight percentile and the total behavioral scale score, along with a positive correlation between hyperactivity and the emotional and behavioral scales. Learning difficulties were more frequent in early-onset FGR than in late-onset FGR. (4) Conclusions: Our study of behavioral development has demonstrated higher behavioral disability rates in children with FGR at 6 years of age; specifically, high rates of hyperactivity or conduct problems. At the same time, we have proved a negative relationship between the birth weight percentile and the total behavioral scale score.

2.
Methods Mol Biol ; 2588: 157-169, 2023.
Article En | MEDLINE | ID: mdl-36418687

The quantitative polymerase chain reaction (qPCR) is a variant of PCR aimed to detect and quantify a targeted DNA molecule. This is made through the addition of probes labeled with fluorescent molecules that emit fluorescence within each amplification cycle, resulting in fluorescence values proportional to the amount of accumulated PCR product. This chapter presents the detailed procedures for quantification of different periodontal pathogens (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus, Streptococcus oralis, and Fusobacterium spp.) using qPCR. It also includes the description of the most frequent problems encountered, how to solve them, and recommendations to minimize the risks for laboratory staff handling oral samples. In addition, a detailed protocol for multiplex qPCR to detect and quantify Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia is also included.


Aggregatibacter actinomycetemcomitans , Tannerella forsythia , Humans , Real-Time Polymerase Chain Reaction , Porphyromonas gingivalis/genetics , Coloring Agents
5.
Arch Oral Biol ; 102: 199-204, 2019 Jun.
Article En | MEDLINE | ID: mdl-31075524

OBJECTIVE: To validate a multiplex real time qPCR (m-qPCR) assay for the simultaneous detection and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in subgingival samples, when compared with anaerobic culture. MATERIAL AND METHODS: Subgingival plaque samples were obtained from patients seeking periodontal treatment. Samples were processed in parallel by anaerobic culturing and by m-qPCR directed to the target bacterial species. Counts and frequency of detection were calculated and analyzed by Mann-Whitney U and chi-square tests, respectively. Contingency tables were constructed, and sensitivity, specificity, predictive values and Lin's correlation coefficients were calculated. RESULTS: Fifty-nine samples were included in the study. A good concordance was achieved between m-qPCR and culture for A. actinomycetemcomitans and P. gingivalis (net agreement, 94.92% and 91.53%, respectively). For T. forsythia, m-qPCR showed statistically significant higher counts than culture (p < 0.005), and low specificity (3.12%) and concordance (47.46%). High sensitivity (above 96.22%) was attained for the three target bacteria with m-qPCR. CONCLUSION: Compared to culture, the tested m-qPCR assay for subgingival plaque samples showed high degree of sensitivity in the simultaneous quantification of A. actinomycetemcomitans, P. gingivalis and T. forsythia.


Dental Plaque , Aggregatibacter actinomycetemcomitans , Anaerobiosis , Humans , Multiplex Polymerase Chain Reaction , Porphyromonas gingivalis , Tannerella forsythia , Treponema denticola
6.
Methods Mol Biol ; 1537: 191-202, 2017.
Article En | MEDLINE | ID: mdl-27924595

The quantitative polymerase chain reaction (qPCR) is a variant of PCR aimed to detect and quantify a targeted DNA molecule through the addition of probes labeled with fluorescent molecules that emit fluorescence within each amplification cycle, what results in fluorescence values proportional to the amount of accumulated PCR product. This chapter presents the detailed procedures for quantification of different periodontal pathogens (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus, and Fusobacterium spp.) using qPCR. It also includes the description of the most frequent problems encountered and how to solve them. In addition, a detailed protocol for multiplex qPCR to detect and quantify P. gingivalis and A. actinomycetemcomitans is included.


Metagenome , Metagenomics , Periodontal Diseases/microbiology , Real-Time Polymerase Chain Reaction , Gingival Crevicular Fluid/microbiology , Humans , Metagenomics/methods , Real-Time Polymerase Chain Reaction/methods
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