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At least 181 records · Page 10Linked to original sources
PubMed2005

Development of a molecular screening test for hereditary hearing loss and genetic susceptibility to aminoglycoside toxicity for Chinese population.

OBJECTIVE: To develop a molecular screening test for genetic defects on hearing loss related genes has significant impacts on early identification of hereditary hearing loss and genetic susceptibility to aminoglycoside ototoxicity. Early identification of pre-lingual hearing loss is very important for patient' s language development, academic achievement, and social skill. Two common mutations, the 235delC in GJB2 gene and the mutation A1555G in mitochondrial DNA, are included in the newly developed screening panel for Chinese population. METHODS: A molecular genetic assay, based on fluorescent labeled multiplex PCR and automatic DNA fragment analyzing techniques, was developed to detect both mutations simultaneously. RESULTS: This assay was able to detect both mutations from patient's samples, and pooled DNA tests, as well as suitable to detect mutation from the DNA extracted from dried blood spot and buccal swab. CONCLUSION: This assay could be a useful tool for newborn screening and carrier screening for the hereditary hearing loss for the Chinese population.

Aminoglycosides↗
PubMed1998

Molecular diagnostic tests for ascertainment of genotype at the mucopolysaccharidosis type VII locus in dogs.

OBJECTIVE: To develop a molecular diagnostic test to ascertain genotype of the mucopolysaccharidosis type VII (MPS VII) locus. SAMPLE POPULATION: Blood samples from 45 mixed-breed (German Shepherd Dog X Beagle) dogs that were phenotypically normal or affected with MPSVII. PROCEDURE: The canine beta-glucuronidase gene (exon 3) was amplified by polymerase chain reaction (PCR), using 2 pairs of primers to determine the genotype of the MPSVII locus by 2 independent methods. For the first method, PCR products were used for heteroduplex analysis, using conformation-sensitive gel electrophoresis. In the second method, an allele-specific restriction site was created by use of a mismatch primer in PCR. The amplified DNA fragment was digested with a restriction enzyme (Eag I) to enable identification of the wild-type and mutant alleles. RESULTS: Conformation-sensitive gel electrophoresis resulted in a single DNA band representing homoduplex when the sample contained a wild-type or MPS VII allele, but 2 bands representing hetero- and homoduplexes when both alleles were in the sample. Restriction digestion of the DNA fragment obtained by use of a mismatch primer was cleaved only when the template was a wild-type allele. Thus, samples from phenotypically normal carrier dogs that contained wild-type and MPS VII alleles were partially digested by the enzyme. CONCLUSIONS: The diagnostic test used 2 strategies for independently ascertaining the wild-type or mutant MPS VII alleles in dogs. Thus, test results can distinguish phenotypically normal MPS VII-carrier dogs from homozygous normal dogs.

Alleles↗
PubMed2006

Multiplex detection of Ehrlichia and Anaplasma pathogens in vertebrate and tick hosts by real-time RT-PCR.

Tick-borne rickettsial infections are responsible for many emerging diseases in humans and several vertebrates. These include human infections with Ehrlichia chaffeensis, Ehrlichia ewingii and Anaplasma phagocytophilum. As single or co-infections can result from a tick bite, the availability of a rapid, multiplex molecular test will be valuable for timely diagnosis and treatment. We recently described a muliplex-molecular test that can detect single or co-infections with up to five Ehrlichia and Anaplasma species. We reported that the test has the sensitivity to identify single infections in the canine host with E. chaffeensis, E. canis, E. ewingii, A. phagocytophilum, and A. platys and co-infection with E. canis and A. platys. In this study, ticks were collected from different parts of the state of Kansas during summer months of the year 2003 and tested for the presence of infection using the molecular test. The analysis revealed a minimum of 3.66% of the ticks to be positive for either E. chaffeensis or E. ewingii in A. americanum and Dermacenter species. This assay will be valuable in monitoring infections in dogs and ticks, and with minor modifications it can be used for diagnosing infections in people and other vertebrates.

Anaplasma↗
PubMed1997

New molecular diagnostic tests for two congenital forms of anemia.

Congenital hypoplastic anemias are a rare and heterogeneous group of disorders. This paper reviews new molecular diagnostic tests in two distinct forms of congenital anemias, anemia due to transplacental infection with B19 parvovirus and Fanconi anemia. In both instances, molecular assays making use of amplification of DNA by the polymerase chain reaction have been used to diagnose either a specific viral infection or gene mutation responsible for a disorder. Recognition of these entities has important prognostic and therapeutic implications.

Anemia↗
PubMed2003

Behavior genetics of canine aggression: behavioral phenotyping of golden retrievers by means of an aggression test.

Molecular genetic analysis of complex traits such as aggression strongly depends on careful phenotyping of individuals. When studying canine aggression, the information provided by the owners of the dogs is often not detailed and reliable enough for this purpose. Therefore we subjected 83 golden retrievers, both aggressive and nonaggressive individuals, to a behavioral test. These tests were analyzed with help of an ethogram, resulting in a behavioral profile for each of the dogs. In this article three methods are described of converting these profiles into a measure of behavioral phenotype. The usefulness of the methods is evaluated by comparing the test results with information provided by owners. Moreover, the hypothesis underlying all these methods, that a lowered threshold for aggressive behavior in general is present in the dogs, is also evaluated. Future research will need to reveal whether the methods meet the high standards that are necessary for studying complex traits.

Aggression↗
PubMed2001

Role of molecular diagnostic testing in familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer families.

PURPOSE: Genetic tests are available for familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer. The goal of this review was to develop an algorithm for application of molecular diagnostic techniques to the management of hereditary colorectal carcinoma and to familiarize the clinician with the vocabulary of molecular genetic testing for hereditary colorectal carcinoma. METHODS: Studies examining the clinical use of genetic testing for hereditary colorectal carcinoma syndromes are evaluated. Recent advances in molecular genetic technology are reviewed, and clinical management as practiced here and elsewhere is outlined. RESULTS: This review is a guide to the most reliable molecular diagnostic techniques. Three key questions are answered: who, when, and how to test. CONCLUSIONS: When integrated with existing testing protocols for colorectal carcinoma and when applied with appropriate caveats, particularly regarding interpretation of negative results, genetic testing can result in improved management of patients and families.

Adenomatous Polyposis Coli↗
PubMed2000

Molecular genetic testing in pediatric practice: A subject review. Committee on Genetics.

Although many types of diagnostic and carrier testing for genetic disorders have been available for decades, the use of molecular methods is a relatively recent phenomenon. Such testing has expanded the range of disorders that can be diagnosed and has enhanced the ability of clinicians to provide accurate prognostic information and institute appropriate health supervision measures. However, the proper application of these tests may be difficult because of their scientific complexity and the potential for negative, sometimes unexpected, consequences for many patients. The purposes of this subject review are to provide background information on molecular genetic tests, to describe specific testing modalities, and to discuss some of the benefits and risks specific to the pediatric population. It is likely that pediatricians will use these testing methods increasingly for their patients and will need to evaluate critically their diagnostic and prognostic implications.

Adolescent↗
PubMed2007

Human papillomavirus testing and molecular markers of cervical dysplasia and carcinoma.

Cervical cancer is the second most common cancer in women worldwide. Human papillomavirus (HPV) is the etiologic agent for the vast majority of premalignant and malignant lesions, and high-risk HPV types can be detected in almost all cases of cervical dysplasia and carcinoma. HPV testing has been widely adopted for the triage of patients after a cervical cytology screening test (Papanicolaou smear or liquid-based cervical cytology such as ThinPrep or SurePath) interpretation of atypical squamous cells of undetermined significance (ASCUS), and HPV testing is increasingly used for screening in conjunction with cervical cytology. Although cervical cytology is a highly effective screening test for cancer, it has limited specificity for clinically significant lesions in cases with low-grade cytologic abnormalities. Up to a quarter of all patients may have a false-negative result on the basis of cervical cytology testing alone. This review focuses on HPV testing methods and molecular markers and their clinical relevance. HPV testing and surrogate molecular markers of HPV infection (p16INK4a) may help identify cases that are associated with underlying high-grade premalignant or malignant lesions and may also reduce aggressive treatment of patients with low-grade lesions.

Alphapapillomavirus↗
PubMed2000

[Quality assurance of the molecular diagnostic tests in hematological malignancies].

Analyses of nucleic acid sequences in hematological malignancies are now essential in managing patients with hematological malignancies. Because of the impact of the test on decision-making in patient care, its accuracy is quite important. Detection of BCR-ABL mRNA by polymerase chain reaction, one of the most popular molecular diagnostic tests for hematological malignancies, has been used for diagnosis of Philadelphia-positive leukemias. Quality of the test must be assured in the process of diagnostic plan, sampling, measurement(nucleic acid isolation, amplification, detection), report and interpretation. Risk of false-positive results due to contamination of in vitro nucleic acid amplification reactions can be decreased through the use of protocols for contamination control. False-negative results may be caused by variability in BCR breakpoint, and loss or degradation of RNA due to inappropriate procedures as well as an insufficient detection sensitivity. Presence of inhibitors of amplification is also to be considered as a cause of false-negative results. Thus, quality control techniques such as internal standards are mandatory to ensure efficient amplification as well as RNA extraction. For the precise evaluation of minimal residual diseases, accurate and sensitive quantitative analyses are required. For quality assurance of assays in the present status of no commercially available kits, it is particularly important to monitor the clinical validation of the results by correlating them with the patient's status to prove clinically relevant. Staffs need to be trained to be familiar with both molecular pathogenesis and technology so that they can provide informative test results with a high quality.

Genes, abl↗
PubMed2002

Apolipoprotein E and other cerebrospinal fluid proteins differentiate ante mortem variant Creutzfeldt-Jakob disease from ante mortem sporadic Creutzfeldt-Jakob disease.

The ability to perform an ante mortem differential diagnosis of Creutzfeld-Jakob disease (CJD) is aided by several clinical and molecular tests. There is a need for molecular tests which can reliably distinguish ante mortem variant CJD (vCJD) from ante mortem sporadic CJD (spCJD). A proteomics approach employing two-dimensional protein electrophoresis is applied to the study of ante mortem CSF samples obtained in collaboration with the CJD Surveillance Unit and the National Hospital for Neurology and Neurosurgery. The sample set includes two cases of vCJD, three cases of spCJD and three neurologic controls. Preliminary data using a panel of seven molecular markers is able to distinguish vCJD from spCJD using a heuristic clustering algorithm. One of the molecular markers has been identified as apolipoprotein E which appears to be upregulated in the cerebrospiral fluid (CSF) of patients with vCJD as compared to spCJD. Analysis of ante mortem CSF may help to differentiate patients with vCJD from those patients with spCJD.

Adult↗
PubMed2001

Canine molecular genetic testing.

Inherited diseases are common among dogs. Recent advances in molecular genetics provide the groundwork for the development of genetic tests for the diagnosis and prevention of inherited diseases. As a result of this progress, genetics should become an integral part of veterinary medicine. DNA tests are safe, easy to perform, and reliable if interpreted correctly. Genetic tests only need to be performed once in a dog's lifetime, because the results of DNA testing never change. Veterinarians should be prepared to understand genetic testing and counseling because they are becoming increasingly important to veterinary medicine.

Animals↗
PubMed2005

Understanding the genotype of follicular thyroid tumors.

Tumors of the thyroid with a follicular growth pattern are controversial and can be diagnostically challenging for the pathologist. This group of tumors includes both follicular derived lesions (adenomas and carcinomas) and papillary carcinoma (follicular variant of papillary carcinoma). H&E morphology has classically been the gold standard for diagnosis. In the past several decades, however, several important molecular markers have been identified that may be unique to different types of thyroid carcinomas. These include the translocations RET/PTC and PAX8-PPARgamma and point mutations in the BRAF and RAS genes. Other molecular events in tumor suppressor genes may be useful for diagnosis of these tumors as well. None of the mutational markers are very sensitive, and there is some question regarding specificity for malignancy, because mutations have also been described in histologically benign tumors. However, with increasing availability of molecular testing for the general pathologist, a molecular testing panel used in conjunction with the H&E morphology and immunohistochemical stains may become useful in the clinical setting for the diagnosis of thyroid tumors.

Carcinoma, Papillary, Follicular↗
PubMed2004

Screening selected blood donors with biochemical iron overload for hemochromatosis: a regional experience.

BACKGROUND AND OBJECTIVES: Hemochromatosis is a genetic disorder characterized by progressive iron overload which leads to early abnormalities of iron parameters (increased transferrin saturation =TS and serum ferritin=SF) and late clinical complications. The disease is prevalently due to C282Y and H63D mutations in the HFE gene, but additional molecular defects are present in a minority of patients. DESIGN AND METHODS: From January to December 2002 we screened first time blood donors of Piedmont, a region of North-western Italy, for TS>45%. Individuals with TS>45% underwent a second fasting check, SF assessment and molecular tests, investigating 12 hemochromatosis-associated molecular defects. RESULTS: A total of 13,998 subjects were screened; 868 (6.2%) had TS>45% and were recalled. Four hundred and eight-six underwent molecular testing. In this selected population allele frequencies of C282Y, H63D and S65C were 6.8%, 22.4% and 1.0%, respectively. No rare mutations were detected, except E168Q in HFE. When measured during fasting, TS was significantly higher in C282Y homozygotes and H63D/C282Y heterozygotes (p<0.05) than in wild type subjects, but not in H63D homozygotes. Hyperferritinemia was documented in 32 cases, 9 with wild type genotype. Mean age, body mass index (BMI) and alcohol intake were higher in this group than in individuals with normal SF. INTERPRETATION AND CONCLUSIONS: This study is an example of a large, two-step hemochromatosis screening with moderate effort and low cost, that enriches basal C282Y allele frequency by about three-fold. Screening based on genotyping only subjects found to have a TS>45% is feasible but, in order to be cost effective should be based on the identification of the two prevalent mutations even in an area where several forms of hemochromatosis have been reported.

Adult↗
PubMed1999

Clinical use of hepatitis C virus tests for diagnosis and monitoring during therapy.

This article reviewed various methods used for the diagnosis and monitoring of HCV infection and discusses potential clinical applications. Substantial improvements have recently been made in assay technology. Moreover, the role of molecular testing in the clinical setting of hepatitis C is becoming better defined. The major challenge facing clinical laboratories is the further refinement, implementation, and standardization of optimized molecular tests, so that reliable data may be made available to clinicians. In turn, clinicians must understand the limitations of each methodology, including the variability of testing that may occur among different laboratories. As more experience is gathered, molecular testing will probably provide important data regarding the most effective use of current and future therapies for individual patients to achieve maximum benefit in the management of hepatitis C.

Hepatitis C↗
PubMed2000

Conventional versus molecular diagnostic tests.

In this session, emphasis was placed on the diagnosis of various mycoses through the identification of antibodies and antigens in sera, as well as on new techniques to properly identify medically important fungi through molecular biological procedures. The use of restriction fragment length polymorphism (RFLP) on fungal mitochondrial DNA (mtDNA) has enabled the identification of different strains of Sporothrix schenkii, several dermatophytes, Candida spp. and black fungi according to their species-specific mtDNA-RFLP patterns. In some species, distinct specific types where found in relation to the geographic origin of the patients. These particular molecular diagnostic tests are useful in the identification of strains and in epidemiological studies. An account of the applications of serological methods in the diagnosis of paracoccidioidomycosis was presented. Serology has been used in the identification of paracoccidioidomycosis using a specific, sensitive and rapid antibody-based immunodiagnosis method. Using the gp43 antigen, the diagnostic coverage of inmunodifussion has been improved from the 93-95% achieved with crude antigens, to 100% in an enzyme-linked immunodiffusion assay capture test. Cross-reactions were eliminated by treatment of the antigen with sodium metaperiodate. Antibody detection is useful, especially in paracoccidioidomycosis and histoplasmosis.

Genetic Techniques↗
PubMed2026

Molecular diagnostic tests for isoniazid-resistant tuberculosis: a scoping review.

The paucity of diagnostic tests for isoniazid-resistant tuberculosis is concerning, given its status as the most common form of drug-resistant tuberculosis and a gateway to multidrug-resistant diseases. Molecular drug-susceptibility testing has improved access to timely diagnosis of rifampicin-resistant tuberculosis, but testing for isoniazid-resistant tuberculosis still remains rare. In this Review, we assessed the characteristics of molecular drug-susceptibility testing for detection of isoniazid-resistant tuberculosis, referencing the WHO target product profiles. 9243 citations were screened to select 238 studies published between 2000 and 2024. The diagnostics options have expanded rapidly since 2020, with 27 nucleic acid amplification tests, eight line probe assays, five DNA microarrays, two targeted next-generation sequencing platforms, and two whole-genome sequencing platforms. Most of the evaluated molecular drug-susceptibility tests met diagnostic performance targets but were often complex and costly. Although a few low-complexity nucleic acid amplification tests met key target product profile criteria, additional field validation and greater efforts are needed to ensure optimal feasibility and affordability for low-resource settings.

Isoniazid↗
PubMed2003

Molecular diagnosis of Prader-Willi syndrome.

BACKGROUND: Prader-Willi syndrome (PWS) is characterized by neonatal hypotonia and feeding problems in infancy, developmental delay, hyperphagia with obesity, short stature, hypogonadism, characteristic facial appearance, and behavior problems. The diagnosis of PWS is based on clinical findings that change with age. PWS has proved to be a difficult condition to recognize with the diagnosis often being delayed until later childhood or even adulthood. Therefore, a molecular testing for PWS is needed to confirm the diagnosis. OBJECTIVE: To study the clinical features of Prader-Willi syndrome patients and confirm diagnosis by molecular testing. MATERIAL AND METHOD: Eighteen Prader-Willi syndrome patients who were diagnosed between March, 1997 and February, 2002 at the Genetic Unit, Queen Sirikit National Institute of Child Health, Bangkok. Peripheral blood lymphocytes were obtained and cultured using the standard technique for chromosome analysis. For fluorescence in situ hybridization (FISH) studies, the specific DNA probes for loci small nuclear ribonucleoprotein polypeptide N (SNRPN) were used to detect deletion. Non-deleted cases were confirmed to have PWS by methylation analysis. RESULTS: The diagnosis of eighteen PWS patients was confirmed by FISH using DNA probes for loci SNRPN demonstrating a deletion of chromosome 15q11-q13 in fourteen cases (77%). Four cases (23%) were confirmed to have PWS resulting from maternal uniparental disomy by demonstrating exclusively maternal specific DNA methylation patterns. CONCLUSION: The clinical diagnosis of PWS should be confirmed by molecular testing especially in the infancy period to avoid needless invasive diagnostic testing.

Child↗