ABSTRACT
Orthoptic and ophthalmologic consultation is an essential step in the diagnosis and treatment of learning disorders, particularly in children with dyspraxia. Such a specialized consultation allows identification of cognitive visual disorders, especially oculomotor or visual-spatial impairment, which disrupt the cognitive processes involved in key academic tasks such as reading or handwriting. It is therefore essential to screen and manage these disorders early in order to implement a rehabilitation plan and counsel educators so as to facilitate learning. After describing cognitive visual disorders in the context of dyspraxia, we discuss principal signs, assessment and ophthalmologic and orthoptic management as conducted in our practice. Several clinical cases illustrate our approach.
Subject(s)
Apraxias/complications , Diagnostic Techniques, Ophthalmological , Ocular Motility Disorders/complications , Ocular Motility Disorders/diagnosis , Vision Disorders/complications , Vision Disorders/diagnosis , Child , Female , Humans , Male , Ophthalmology , OrthopticsABSTRACT
Diagnosis of central nervous system (CNS) infection with herpes simplex virus (HSV) requires sensitive and rapid techniques. PCR therefore is considered to be the diagnostic gold standard in these cases. However, current PCR protocols are time-consuming and labor-intensive. In addition, the need for post-amplification manipulations increases the risk of laboratory contaminations with amplified products. In order to improve conventional PCR techniques we compared our current semiautomated HSV-PCR-ELISA assay with a new micro-volume rapid-cycle PCR system that combines real-time monitoring and fluorescence melting-curve analysis without the need for post-amplification sample manipulations. Spiking experiments with supernatants of tissue culture-grown HSV type 1 (HSV-1) and type 2 (HSV-2) in HSV-negative control cerebrospinal fluid (CSF) and sterile water revealed that the new rapid cycle PCR protocol is as sensitive and specific as the PCR-ELISA. Furthermore, a mismatch (G:T) within the probe-targeted region of the HSV-2 glycoprotein B gene decreases the probe/product melting temperature (Tm) from 69 degrees C for HSV-1 to 64 degrees C for HSV-2, enabling the simultaneous identification of the two HSV genotypes by melting-curve analysis within one run. This type specificity of the system was confirmed with 30 genital swabs previously analyzed for the presence of HSV-1/2 in cell culture. While our current PCR-ELISA method needs up to 1 day from sample preparation to result generation, the new procedure takes only 1 h. We consider this system as a promising new tool for the analysis of HSV DNA in CSF and in other human body fluids as well as for the diagnosis of other infectious agents where rapid diagnosis, high sensitivity and specificity are required.
Subject(s)
Central Nervous System Infections/virology , DNA, Viral/cerebrospinal fluid , Herpes Simplex/diagnosis , Herpesvirus 1, Human/genetics , Herpesvirus 2, Human/genetics , Central Nervous System Infections/cerebrospinal fluid , DNA Primers , Fluorescence , Genotype , Herpes Simplex/cerebrospinal fluid , Herpes Simplex/genetics , Herpesvirus 1, Human/isolation & purification , Herpesvirus 2, Human/isolation & purification , Humans , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Sensitivity and SpecificityABSTRACT
Varicella syndrome (VS) specific malformations were sonographically seen at 22 weeks and 3 days of gestation. Fetal infection was demonstrated by detection of varicella-zoster virus (VZV) DNA in fetal blood and amniotic fluid by polymerase chain reaction (PCR). Following therapeutic abortion, fetal infection was confirmed by detection of VZV DNA in several fetal tissues and placenta, and by histopathological findings like miliary calcified necroses in fetal organs.
Subject(s)
Chickenpox/transmission , Fetal Diseases/diagnostic imaging , Herpesvirus 3, Human/isolation & purification , Infectious Disease Transmission, Vertical , Pregnancy Complications, Infectious , Prenatal Diagnosis , Abortion, Therapeutic , Adult , Amniotic Fluid/virology , Chickenpox/diagnostic imaging , Chickenpox/embryology , DNA, Viral/isolation & purification , Female , Fetal Diseases/embryology , Herpesvirus 3, Human/genetics , Humans , Pregnancy , Syndrome , UltrasonographyABSTRACT
Up to now 19 allelic sequences of the rhesus monkey DQB1 locus have been published. Referring to these sequences, we have developed a typing protocol for Mamu-DQB1 alleles which was verified by additional cloning, sequence analysis and segregation studies. The protocol is based on the amplification of the second exon with only one specific primer pair followed by the digestion of the PCR products with up to 10 different restriction endonucleases. The alleles can be identified in homozygous and heterozygous combinations since most amplified second exon sequences give unique hand patterns after digestion with at least one of the selected restriction endonucleases. By the use of this protocol we analyzed DNA-samples from 182 rhesus monkeys. Among these samples two novel Mamu-DQB1 alleles were detected, subsequently cloned and their nucleic sequence determined. Since we typed four complete breeding groups consisting of two generations we were able to identify several DQ haplotypes by segregation analysis using the previously developed typing protocol for DQA1.
