Rapid detection of high-risk HPV16 and HPV18 based on microchip electrophoresis / 药物分析学报

Researches on detection of human papillomavirus (HPV) high-risk samples were carried out by poly-merase chain reaction (PCR) coupled with microchip electrophoresis (MCE). Herein, we introduced a simple, rapid, automated method for detecting high-risk samples HPV16 and HPV18. In this research, general primers were initially selected to obtain sufficient detectable yield by PCR to verify feasibility of MCM method for HPV detection, then type-specific primers were further used to evaluate the specificity of MCE method. The results indicated MCE method was capable of specifically detecting high-risk HPV16 and HPV18, and also enabled simultaneous detection of multiplex samples. This MCE method described here has been successfully applied to HPV detection and displayed excellent reliability demonstrating by sequencing results. The inherent capability of MCE facilitated HPV detection conducted in a small chip with automated, high throughput, massive parallelized analysis. We envision that MCE method will definitely pave a way for clinical diagnosis, and even on-site screening of cervical cancer.

Diagnóstico molecular de tuberculosis multidrogorresistente en muestras de esputo mediante el análisis de curvas de melting / Molecular diagnosis of multidrug-resistant tuberculosis in sputum samples by melting curve analysis

RESUMEN Objetivos. Analizar curvas de melting para el diagnóstico de tuberculosis multidrogorresistente a partir de muestras de esputo. Materiales y métodos. Se colectaron muestras de esputo (n = 250) de pacientes con sospecha clínica de tuberculosis pulmonar según resultado de baciloscopia y cultivados en medio sólido Lowenstein Jensen. Según el método de referencia se trabajó con 124 muestras sensibles a rifampicina e isoniacida, 24 resistentes a rifampicina, 33 resistentes a isoniacida y 69 multidrogorresistentes. Se evaluó por PCR en tiempo real y luego por las curvas de melting, se utilizó el gen rpoB como biomarcador de resistencia a rifampicina, y el gen katG y región promotora inhA como biomarcadores de resistencia a isoniacida. La cepa H37Rv fue considerada como control sensible a drogas. Se compararon los resultados del método de referencia y los resultados del análisis de curvas de melting para evaluar los parámetros de sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo. Resultados. La resistencia a rifampicina mostró una sensibilidad de 90,3 %, especificidad de 90,4 %, valor predictivo positivo de 84,8 % y valor predictivo negativo de 94,0 %. La resistencia a isoniacida mostró una sensibilidad de 90,2 %, especificidad de 93,9 %, valor predictivo positivo de 91,1 % y valor predictivo negativo de 93,3 %. La detección de tuberculosis multidrogorresistente mostró valores de 89,9 %, 90,6 %, 78,5 % y 95,9 % para sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo, respectivamente. Conclusiones. El análisis de curvas de melting mostró ser seguro y confiable para ser utilizado en el diagnóstico rápido de tuberculosis multidrogorresistente en muestras de esputo.

ABSTRACT Objectives. To analyze melting curves for the diagnosis of multidrug-resistant tuberculosis from sputum samples. Materials and Methods. Sputum samples (n = 250) were collected from patients with clinical suspicion of pulmonary tuberculosis as a result of bacilloscopy and cultured in solid medium Lowenstein Jensen. According to the reference method, 124 samples sensitive to rifampicin and isoniazid, 24 resistant to rifampicin, 33 resistant to isoniazid, and 69 multidrug-resistant were used. It was evaluated by real-time PCR and then by melting curves, the rpoB gene was used as a biomarker of rifampicin resistance, and the katG gene and inhA promoter region were used as biomarkers of isoniazid resistance. The H37Rv strain was considered a drug-sensitive control. The results of the reference method and the results of the melting curve analysis were compared to evaluate the parameters of sensitivity, specificity, positive predictive value and negative predictive value. Results. Rifampicin resistance showed a sensitivity of 90.3%, specificity of 90.4%, positive predictive value of 84.8% and negative predictive value of 94.0%. Isoniazid resistance showed a sensitivity of 90.2%, specificity of 93.9%, positive predictive value of 91.1% and negative predictive value of 93.3%. The detection of multidrug-resistant tuberculosis showed values of 89.9%, 90.6%, 78.5% and 95.9% for sensitivity, specificity, positive predictive value and negative predictive value, respectively. Conclusions. The melting curve analysis showed to be safe and reliable to be used in the rapid diagnosis of multidrug-resistant tuberculosis in sputum samples.

Prevalence of Complement-Mediated Cell Lysis-like Gene (sicG) in Streptococcus dysgalactiae subsp. equisimilis Isolates From Japan (2014–2016)

BACKGROUND: Streptococcus dysgalactiae subsp. equisimilis (SDSE; a β-hemolytic streptococcus of human or animal origin) infections are emerging worldwide. We evaluated the clonal distribution of complement-mediated cell lysis-like gene (sicG) among SDSE isolates from three central prefectures of Japan. METHODS: Group G/C β-hemolytic streptococci were collected from three institutions from April 2014 to March 2016. Fifty-five strains (52 from humans and three from animals) were identified as SDSE on the basis of 16S rRNA sequencing data.; they were obtained from 25 sterile (blood, joint fluid, and cerebrospinal fluid) and 30 non-sterile (skin-, respiratory tract-, and genitourinary tract-origin) samples. emm genotyping, multilocus sequence typing, sicG amplification/sequencing, and random amplified polymorphic DNA (RAPD) analysis of sicG-positive strains were performed. RESULTS: sicG was detected in 30.9% of the isolates (16 human and one canine) and the genes from the 16 human samples (blood, 10; open pus, 3; sputum, 2; throat swab, 1) and one canine sample (open pus) showed the same sequence pattern. All sicG-harboring isolates belonged to clonal complex (CC) 17, and the most prevalent emm type was stG6792 (82.4%). There was a significant association between sicG presence and the development of skin/soft tissue infections. CC17 isolates with sicG could be divided into three subtypes by RAPD analysis. CONCLUSIONS: CC17 SDSE harboring sicG might have spread into three closely-related prefectures in central Japan during 2014–2016. Clonal analysis of isolates from other areas might be needed to monitor potentially virulent strains in humans and animals.

Analysis of bite mark evidence.

Bite mark analysis casework strives to connect a biter to the teeth pattern present on an object linked in some way to a crime or event. The general public and some law enforcement may consider any “bite mark” case they develop to be a certainty in the quest to identify the biter. The ability of skin to register sufficient detail of a biter’s teeth is highly variable. Bite mark casework indicates that many bite marks are not well defined in detail and posses distortion due to the physical nature of skin itself. The current opinion is that bite mark can be useful in including or excluding possible suspects and ability to identify only one person as the biter. In mortal combat situations, such as the violence associated with life and death struggles between assailants and victims, the teeth are often used as a weapon. It is well known that assailants in sexual attacks, including sexual homicide, rape and child sexual abuse, often bite their victims as an expression of dominance, rage and animalistic behaviour. The teeth are a significant component of our natural arsenal.

Two Human Cases Infected by the Horsehair Worm, Parachordodes sp. (Nematomorpha: Chordodidae), in Japan

The present study was performed to describe 2 human cases infected by the horsehair worm, Parachordodes sp., in Japan. Two gordiid worms were collected in the vomit and excreta of an 80-year-old woman in November 2009 in Kyoto city, and in the mouth of 1-year-old boy in December 2009 in Nara city, Japan, respectively. Both worms were males having bifurcated posterior ends and male gonads in cross sectional specimens. They were identified as Parachordodes sp. (Nematomorpha: Chordodidae) based on the characteristic morphologies of cross sections and areoles in the cuticle. DNA analysis on 18S rRNA partial sequence arrangements was also carried out and both worms were assumed to be close to the genus Paragordionus based on tree analysis, and far from Gordius sp. which has already been reported in humans in Japan. DNA sequencing of the Parachordodes worm does not appear on the database; therefore, more information on the gene sequences of the genus Parachordodes from humans, animals, or intermediates is required.

An approach for identification of individuals in a mass disaster in Indian set up.

In the changing scenario of mass disaster, which has become almost one of the stories of daily news paper, Disaster Victim Identification has become important for its medico-legal as well as nations socio-economical aspects. This expertise gives the Forensic Experts a resource advice in dealing with human identification in a simpler way in Indian context. The idea of this work was born after the Ganeswari express accident that occurred in West Bengal, in May 2010, after which the situation demeaned a discussion between the Forensic Experts, Forensic Scientists and police personnel to frame a simpler guideline for Disaster Victim Identification. The recommendations made in this paper, as well as in many cited references, are intended to provide the forensic experts the minimum guidance for victim identification by photography and storing samples suitable for DNA analysis. It also gives a guideline for matching the DNA samples with the relative of the victims that will provide better chances of victim identification.

Quantitative DNA analysis in diagnosis of benign and malignant breast masses / 中国综合临床

Objective To explore the application of quantitative DNA analysis in differential diagnosis of benign and malignant breast masses to aid the surgery plan formation.Methods Four hundred and eighty-eight patients with breast mass were enrolled into this study.Tissues of breast mass in patients were gained by fine-needle aspiration puncture.Two sections were made from each sample,one was stained by Papanicolaou for regular cytology analysis and another was stained with Feulgen for quantitative DNA analysis.Pathological results were confirmed in each case after surgery.Results One hundred and sixty-four cases were classified as patients with benign neoplasm,while the other 324 cases were classified as malignant neoplasm,according to the pathological examination results.The sensibility and specificity were 91.4%(296/324) and 92.7%(152/164) for regular cytological method,90.1%(292/324) and 100.0%(164/164) for quantitative DNA analysis method.Meanwhile the positive predictive and negtive value of quantitative DNA analysis was 100.0%(292/292) and 83.7%(164/164),of which regular cytological methods were 96.1%(296/308) and 84.4%(152/180).Conclusion The quantitative DNA analysis might assistant differential diagnosis of benign and malignant breast tumor.

Variabilidade genética na região its do rDNA de isolados de trichoderma spp. (Biocontrolador) e Fusarium oxysporum f. sp. Chrysanthemi / Genetic variability in rDNA ITS region of Trichoderma spp. (biocontrole agent) and Fusarium oxysporum f. sp. chrysanthemi isolates

A análise de características morfológicas e culturais podem não ser suficientes para uma caracterização precisa das espécies de Trichoderma e Fusarium. Objetivou-se, neste trabalho, caracterizar a região do Espaço Interno Transcrito (ITS) do rDNA dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma spp. utilizados no biocontrole de Fusarium oxysporum f. sp. chrysanthemi (isolado) UFSMF6. A extração de DNA de cada isolado foi realizada a partir de micélio produzido em meio líquido Batata-Dextrose. As amostras de DNA genômico foram submetidas à Reação em Cadeia da Polimerase (PCR) com os oligonucleotídeos iniciadores universais ITS1 e ITS4 e o produto gerado foi sequenciado. Os fragmentos gerados pela amplificação da PCR foram tratados com as enzimas de restrição HaeIII, HinfI e MboI. As regiões ITS1, ITS2 e 5.8S do rDNA desses isolados fúngicos foram amplificadas com sucesso. A região ITS dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma e o isolado UFSMF6 de Fusarium apresentaram uma banda simples com um fragmento de aproximadamente 600 pares de base (pb). As enzimas de restrição HaeIII, HinfI e MboI geraram polimorfismo de bandas entre os isolados. Com base nas análises da sequência de DNA, os isolados UFSMT15.1, UFSMT16, UFSMT17 e UFSMF6 apresentaram maior similaridade com as espécies Trichoderma koningiopsis, Hypocrea virens, Hypocrea lixii e Fusarium oxysporum, respectivamente.

The analysis of morphological and cultural characteristics may not enough for the characterization of the species of Trichoderma and Fusarium. The aim of this work was to characterize the Internal Transcribed Spacer (ITS) region of the rDNA of UFSMT15.1, UFSMT16 and UFSMT17 isolates of Trichoderma spp. used in the biocontrol of Fusarium oxysporum f. sp. chrysanthemi UFSMF6. DNA extraction of each isolate was accomplished starting from hyphae produced in liquid medium Potato-Dextrose-Agar. The samples of genomic DNA were submitted to the Polymerase Chain Reaction (PCR) with the primers ITS1 and ITS4, and the product was sequenced. The fragments generated from PCR amplification were digested with the restriction enzymes HaeIII, HinfI and MboI. The ITS region of the Trichoderma's isolates UFSMT15.1, UFSMT16 and UFSMT17 and the Fusarium UFSMF6 isolate showed a simple band with a fragment with 600 base pairs (bp) approximately. The restriction enzymes HaeIII, HinfI and MboI generated band polymorphism among the isolates. The isolates UFSMT15.1, UFSMT16, UFSMT17 and UFSMF6 showed high similarity whit Trichoderma koningiopsis, Hypocrea virens, Hypocrea lixii and Fusarium oxysporum species, respectively.

Living Donor Liver Transplantation in a Korean Child with Glycogen Storage Disease Type IV and a GBE1 Mutation

Glycogen storage disease type IV (GSD-IV) is an autosomal recessive disease caused by a deficient glycogen branching enzyme (GBE), encoded by the GBE1 gene, resulting in the accumulation of abnormal glycogen deposits in the liver and other tissues. We treated a 20-month-old girl who presented with progressive liver cirrhosis and was diagnosed with GSD-IV, as confirmed by GBE1 gene mutation analysis, and underwent living related heterozygous donor liver transplantation. Direct sequencing of the GBE1 gene revealed that the patient was compound heterozygous for a known c.1571G>A (p.Gly264Glu) mutation a novel c.791G> A (Arg524Gln) mutation. This is the first report of a Korean patient with GSD-IV confirmed by mutation analysis, who was treated successfully by liver transplantation.

Significance of Molecular Diagnosis using Histopathological Specimens in Cestode Zoonoses

Cestode zoonosis cases confirmed by PCR-based mitochondrial DNA analysis were investigated. The cestodiosis included taeniasis, cysticercosis, alveolar echinococcosis, cystic echinococcosis, sparganosis mansoni, diphyllobothriasis and diplogonoporiasis. DNA samples were extracted from the ethanol-fixed, formalin-fixed, paraffin-embedded sections, HE-stained, and the PAS- or acetocarmine-stained samples submitted for histopathology. For PCR-based analysis, cytochrome <I>c</I> oxidase subunit 1 and⁄or cytochrome <I>b</I> genes were amplified by multiplex PCR or conventional PCR coupled with DNA sequencing. Although DNA molecules were degraded in most formalin-fixed samples, smaller gene fragments were successfully amplified and the species causing cestodiosis could be identified by DNA sequence analysis of the amplicons. This review describes cestode zoonosis cases in which mitochondrial DNA analysis was useful not only for routine and retrospective diagnosis, but also for genetic polymorphism analysis and molecular identification of the species associated with pathogenicity. The significance of molecular diagnosis using histopathological specimens for cestode zoonoses is also discussed.

Molecular Epidemiological Typing of Methicillin-Resistant Staphylococcus aureus Using Random Amplified Polymorphic DNA Analysis / 감염과화학요법

BACKGROUND: The random amplified polymorphic DNA (RAPD) analysis was investigated to see if this method could be a useful tool for monitoring of epidemic outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) among patients and healthcare workers (HCW) in the intensive care units (ICU). METHODS: Thirty-eight MRSA strains were isolated from patients and HCW in Dong-A University Hospital ICU from October, 1998 to December, 1998 (10 patients and 8 HCW) and May, 2001 to July, 2001 (15 patients and 5 HCW). All strains were typed according to antimicrobial susceptibility and RAPD analysis patterns. mecA genes were detected using polymerase chain reaction (PCR). RESULTS: Twenty one of 25 (84%) and 12 of 13 (92%) MRSA, isolated from patients and HCW, respectively, were mecA positive. mecA positive MRSA were classified into 18 different types by RAPD analysis. CONCLUSION: DNA fingerprinting using RAPD analysis is a simple, effective, and rapid method for discriminating MRSA strains, and may be applicable in detecting outbreaks of S. aureus infections in the ICU.

Molecular Epidemiological Typing of Methicillin-Resistant Staphylococcus aureus Using Random Amplified Polymorphic DNA Analysis / 감염과화학요법

BACKGROUND: The random amplified polymorphic DNA (RAPD) analysis was investigated to see if this method could be a useful tool for monitoring of epidemic outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) among patients and healthcare workers (HCW) in the intensive care units (ICU). METHODS: Thirty-eight MRSA strains were isolated from patients and HCW in Dong-A University Hospital ICU from October, 1998 to December, 1998 (10 patients and 8 HCW) and May, 2001 to July, 2001 (15 patients and 5 HCW). All strains were typed according to antimicrobial susceptibility and RAPD analysis patterns. mecA genes were detected using polymerase chain reaction (PCR). RESULTS: Twenty one of 25 (84%) and 12 of 13 (92%) MRSA, isolated from patients and HCW, respectively, were mecA positive. mecA positive MRSA were classified into 18 different types by RAPD analysis. CONCLUSION: DNA fingerprinting using RAPD analysis is a simple, effective, and rapid method for discriminating MRSA strains, and may be applicable in detecting outbreaks of S. aureus infections in the ICU.

The differential diagnostic values of DNA analysis in malignant and tuberculostic pleural effusion compared with CEA / 中国呼吸与危重监护杂志

Objective To investigate the differential diagnostic values of DNA analysis for malignant and tuberculosis pleural effusion by flow cytometry (FCM), and compare with CEA assay MethodsUsing FCM to detect DNA aneuploidy in malignant and tuberculostic pleural effusion Results Both the specificity and diagnostic index of DNA analysis were better than CEA( P

Molecular typing of Helicobacter pylori by random amplified polymorphic DNA analysis in patients with gastroduodenal diseases / 대한내과학회지

BACKGROUND: Genetic diversity among Helicobacter pylori strains isolated from different patients has been debated. This study was undertaken to determine molecular types and genetic diversity among 20 isolates of H. pylori obtained from various gastroduodenal diseases, and also examined the association between molecular types of H. pylori and these diseases. METHODS: Antral biopsies were taken for culture from 38 patients with chronic gastritis, gastric ulcer and duodenal ulcer at the time of endoscopy. Biopsy specimens were primarily inoculated on chocolate agar and incubated microaerophilically at 37degree Cfor up to 7 days. H. pylori was identified by typical Gram stain morphology and biochemical tests. Random amplified polymorphic DNA (RAPD) fingerprinting was performed by 4 primers (OPA-07, 5'-GAAACGGGTG-3'; OPA-10, 5'-GTGATCGCAG-3'; OPA-11, 5'-CAATCGCCGT-3'; OPA-12, 5'-TCGGCGATAG-3'; Operon Technologies, Atlanta, GA). We used the NTSYS-pc (numerical taxonomy system and multivariate analysis system, version 1.50, Applied Biostatistics Inc., CA, USA) program to compose the phenogram for the differentiation of H. pylori strains. RESULTS: Twenty strains (52.6%) of H. pylori were isolated from 38 biopsy specimens. All isolates were divided into five molecular types (I-V) at similarity (S) value of 0.63; 7 strains (35%), 4 strains (20%), 4 strains (20%), 3 strains (15%) and 2 strains (10%) belonged to type II, III, IV, V and I, respectively. The distribution of genetic S value was 0.24 to 0.91 in all isolates, thus the isolates had a wide range of S values. The mean S values of all isolates, type I, II, III, IV and V were 0.69, 0.69, 0.73, 0.75 and 0.65, respectively. There was no specific correlation between molecular types and gastroduodenal diseases. CONCLUSION: H. pylori isolates had high level of genetic diversity. The RAPD molecular types of H. pylori were not disease-specific since the types were diverse in the isolates from various gastroduodenal diseases.

DNA analysis of hepatocellular carcinoma.

Specimens from 13 cases of resectable hepatocellular carcinoma (HCC) were studied using flow cytometry. The correlation between histological features and ploidy distribution showed those with higher histological grading (HCC grade III or IV) to display an increased tendency towards aneuploidy and polyploidy than those with low grade (HCC grade II and clear cell type of HCC). Along with the tumor’s increased tendency towards aneuploidy and polyploidy, the percent DNA distribution in the S- and G2 – phase was also found increased. The same applied to the DNA index, we also detected a strong correlation between proliferation index and ploidy pattern of HCC.

Molecular Epidemiology of Staphylococcus aureus Isolated in the Hospital Environments / 감염

BACKGROUND: This study was designed to investigate the isolation rate of Staphylococcus aureus from hands and nasal cavities of physicians and nurses and to determine the relationship between the isolates of S. aureus from patients with nosocomial infection and the isolates from physicians, nurses and the hospital environment. METHODS: Twenty-three S. aureus isolates which consisted of 8 strains from patients, 7 strains from doctors and nurses, 7 strains from hospital environments, and S. aureus ATCC 25923 were examined. Biochemical profile by the use of API Staph (API system, La Balme-Les Grottes, France), antibiogram by disk diffusion method, and random amplified polymorphic DNA analysis (RAPD) were used for the strain differentiation and molecular typing of S. aureus isolates. RESULTS: The isolation rate of S. aureus in hands and nasal cavities of doctors and nurses was 21% (25/ 120) and 28% (33/120), respectively. The isolation frequency of methicillin-resistant S. aureus (MRSA) from doctors, nurses, and hospital environments was 57% (8/ 14). The isolates disclosed 13 different biochemical profiles and 14 different resistant patterns of antimicrobials. All isolates were divided into five molecular types (A~E) by RAPD with a similarity (S) value of 0.55; 10 strains (45%) belonged to type A, 7 (32%) to type C, 3 (14%) to type B, one each to type D and E, respectively. The S value between strain 11 from hospital environments and strain 12 from a patient was 1.00, which means that they are the same on the genetic level. CONCLUSION: These findings reveal the existence of a close relationship between clinical isolates and isolates from hospital environment, including those from physicians and nurses. Thus, the implementation of an infection control programs to reduce the spread of MRSA within hospitals is important for the prevention of nosocomial infections.

Molecular Typing of Salmonella typhi by Random Amplified Polymorphic DNA Analysis / 대한임상미생물학회지

BACKGROUND: In the year 1996, there were some outbreaks of Salmonella typhi infection in Pusan and therefore, the incidence of S. typhi infection was markedly increased in comparison with the previous year. To differentiate the isolates epidemiologically, a random amplified polymorphic DNA(RAPD) fingerprinting method has been developed. METHODS: A total of 9 arbitrary primers were screened with S. typhi strains isolated in Pusan, 1996. This allowed selection of a panel of primers capable of detecting DNA polymorphisms among S. typhi isolates. This panel was used to examine 54 strains of S. typhi, which had been isolated in Pusan including the cases of outbreaks that was previously characterized by phage typing. RESULTS: Four single primers and one combination of two primers were selected to discriminate the S. typhi isolates. RAPD analysis resolved the 54 strains into 20 different subtypes. At least two outbreaks were found by RAPD analysis. The isolates of E1 phage type, which are the most common in Korea, were perfectly differentiated with each other, except the strains isolated within the outbreaks. CONCLUSION: The RAPD approach is the useful epidemiologic tool to S. typhi subtyping, which is providing high discriminatory power. There were at least two outbreaks when the epidemic Salmonella infections of Pusan in 1996 had been occurred. The primers or their comb ination capable to discriminate the S. typhi isolates were described.

Clear cell sarcoma of the kidney : immunohistochemical study and flow cytometric DNA analysis of 7 cases

Immunohistochemical study and flow cytometric DNA analysis were done on seven cases of clear cell sarcoma of the kidney (CCSK) to speculate its histogenesis and to access the diagnostic usefulness of these methods in the differential diagnosis of Wilm's tumor. Clinically, CCSK is a rare malignant renal tumor of children with a propensity to metastasize to bone. Arborizing vascular pattern surrounding the tumor cells which have clear cytoplasm is characteristic histologic finding. Immunohistochemically, only vimentin was diffusely demonstrated in the tumor cell membrane and cytoplasm. In flow cytometric DNA analysis, four cases showed diploidy and two cases near diploidy. CCSK is a separate disease entity with characteristic clinicopathologic, immunohistochemical and flow cytometric findings in distinction from Wilms' tumor. Considering the histologic and immunohistochemical findings, the possible histogenetic mechanism of CCSK seems to be in common with congenital mesoblastic nephroma (CMN), that is primitive mesenchymal cells which committed early stromagenic activity.

Comparison of enzyme and DNA analysis in a Tay-Sachs disease carrier screening program

Tay-Sachs disease (GM2 gangliosidosis, type 1; TSD) is an autosomal recessive GM2 gangliosidosis resulting from the deficient activity of the lysosomal hydrolase beta-hexosaminidase A (Hex A). With a carrier frequency estimated at 1 in 25, it is a common lysosomal disorder in the Ashkenazi Jewish population. Tay-Sachs disease has provided the prototype for the prevention of severe recessive genetic diseases. Molecular analysis of the Hex A gene (HEXA) of Ashkenazi Jewish individuals affected with Tay-Sachs disease revealed that three common mutations cause the infantile and adult onset forms of the disease; a four base insertion in exon 11, a splice junction mutation in intron 12 and a point mutation in exon 7 (G269S). A study was undertaken to determine whether mutation analysis would be useful in TSD screening programs in identifying carriers and clarifying the status of individuals whose enzyme assays are inconclusive. Ashkenazi Jewish individuals who had been diagnosed as carriers, inconclusives by enzyme assay and non-carriers with low normal enzyme levels in the Mount Sinai Tay-Sachs Disease Prevention Program were examined for the presence of the three mutations using polymerase chain reaction (PCR) and allele specific oligonucleotide (ASO) hybridization. The insertion mutation was present in 29 of 34 carriers and 2 of 36 inconclusive individuals, the splice junction mutation was found in 4 of 34 carriers and the G269S mutation was found in 1 of 34 carriers. Of the 313 non-carrier individuals with normal enzyme activity in the lower normal range, one was positive for the splice junction mutation.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative study on results between urine flow cytometric DNA analysis and urine cytology in transtional cell carcinoma of bladder / 대한비뇨기과학회지

We compared the roles of urinary cytology and flow cytometric DNA analysis in the evaluation of 26 patients with transitionsl cell carcinoma of bladder from March 1989 to April 1991. When carcinoma was present at the time of specimen collection it was detected by cytology in 65.4 percent and flow cytometric DNA analysis in 73.1 percent. Combination of flow cytemetric DNA analysis and urinary cytology increased the diagnostic yield to 88.5 percent Flow cytometric DNA analysis was slightly more sensitive than urinary cytology for the detection of abnormalities in specimen from low stage. high grade. small size. small number and recurrent cancer but no statistically significant difference was identified. When used in conjunction with urinary cytology. urine flow cytometric DNA analysis was valuable procedure in the diagnosis and follow up of patients with bladder cancer.