RESUMO
Background: asthma is very common in children and its diagnosis is based on clinical manifestations, which can be misdiagnosed as other respiratory diseases with similar signs and symptoms
Objective: to analyze the expression of ST2L and CD203c in the diagnosis of pediatric asthma
Methods: basophils were purified from whole blood samples of patients and healthy controls using Ficol-Paque gradient and Basophil Isolation Kit. RNA extraction was done by RNX-Plus solution and after synthesis of cDNA, the gene expression was analyzed by means of real time PCR
Results: patients expressed significantly higher levels of CD203c than healthy controls [p=0.01]. Although there was an increase in the transcription level of ST2L gene in patients, the results were not statistically significant compared to those obtained from the healthy controls [p>0.05]. A Specificity of 60% and a sensitivity of 73% were foundusing ROC curve for CD203c expression. Patients with positive family history of asthma exhibited more CD203c and ST2L expression [p<0.05]
Conclusion: it is proposed that determining CD203c expression by real time PCR may be an effective technique for diagnosis of pediatric asthma
RESUMO
Background: Maternal-fetal RhD antigen incompatibility causes approximately 50% of clinically significant alloimmunization cases. The routine use of prophylactic anti-D immunoglobulin has dramatically reduced hemolytic disease of the fetus and newborn. Recently, fetal RHD genotyping in RhD negative pregnant women has been suggested for appropriate use of anti-D immunoglobulin antenatal prophylaxis and decrease unnecessary prenatal interventions
Materials and Methods: In this prospective cohort study, in order to develop a reliable and non-invasive method for fetal RHD genotyping, cell free fetal DNA [cffD-NA] was extracted from maternal plasma. Real-time quantitative polymerase chain reaction [qPCR] for detection of RHD exons 7, 5, 10 and intron 4 was performed and the results were compared to the serological results of cord blood cells as the gold standard method. SRY gene and hypermethylated Ras-association domain family member 1 [RASSF1A] gene were used to confirm the presence of fetal DNA in male and female fetuses, respectively
Results: Out of 48 fetuses between 8 and 32 weeks [wks] of gestational age [GA], we correctly diagnosed 45 cases [93.75%] of RHD positive fetuses and 2 cases [4.16%] of the RHD negative one. Exon 7 was amplified in one sample, while three other RHD gene sequences were not detected; the sample was classified as inconclusive, and the RhD serology result after birth showed that the fetus was RhD-negative
Conclusion: Our results showed high accuracy of the qPCR method using cffDNA for fetal RHD genotyping and implicate on the efficiency of this technique to predict the competence of anti-D immunoglobulin administration