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21.
High throughput parallel analysis of hundreds of patient samples for more than 100 mutations in multiple disease genes 总被引:5,自引:0,他引:5
Shuber AP; Michalowsky LA; Nass GS; Skoletsky J; Hire LM; Kotsopoulos SK; Phipps MF; Barberio DM; Klinger KW 《Human molecular genetics》1997,6(3):337-347
As more mutations are identified in genes of known sequence, there is a
crucial need in the areas of medical genetics and genome analysis for
rapid, accurate and cost-effective methods of mutation detection. We have
developed a multiplex allele-specific diagnostic assay (MASDA) for analysis
of large numbers of samples (> 500) simultaneously for a large number of
known mutations (> 100) in a single assay. MASDA utilizes
oligonucleotide hybridization to interrogate DNA sequences. Multiplex DNA
samples are immobilized on a solid support and a single hybridization is
performed with a pool of allele-specific oligonucleotide (ASO) probes. Any
probes complementary to specific mutations present in a given sample are in
effect affinity purified from the pool by the target DNA. Sequence-specific
band patterns (fingerprints), generated by chemical or enzymatic sequencing
of the bound ASO(s), easily identify the specific mutation(s). Using this
design, in a single diagnostic assay, we tested samples for 66 cystic
fibrosis (CF) mutations, 14 beta-thalassemia mutations, two sickle cell
anemia (SCA) mutations, three Tay-Sachs mutations, eight Gaucher mutations,
four mutations in Canavan disease, four mutations in Fanconi anemia, and
five mutations in BRCA1. Each mutation was correctly identified. Finally,
in a blinded study of 106 of these mutations in > 500 patients, all
mutations were properly identified. There were no false positives or false
negatives. The MASDA assay is capable of detecting point mutations as well
as small insertion or deletion mutations. This technology is amenable to
automation and is suitable for immediate utilization for high-throughput
genetic diagnostics in clinical and research laboratories.
相似文献
22.
Oocyte morphology predicts outcome of intracytoplasmic sperm injection 总被引:10,自引:14,他引:10
Serhal PF; Ranieri DM; Kinis A; Marchant S; Davies M; Khadum IM 《Human reproduction (Oxford, England)》1997,12(6):1267-1270
To examine the influence of cytoplasmic morphology on the success rate of
intracytoplasmic sperm injection (ICSI), the morphology of 837 metaphase II
oocytes was assessed after cumulus stripping. The main abnormalities
detected were excessive granularity, cytoplasmic inclusions such as
vacuoles, smooth endoplasmic reticulum clustering and refractile bodies.
Microinjection was performed in 538 oocytes with normal cytoplasm, 142 out
of 161 with excessive granularity and 112 out of 138 with cytoplasmic
inclusions. Very poor oocytes were not injected. No difference was found in
fertilization rate. The embryos achieved cleaved normally and a similar
number of good quality embryos among the three groups was noted. The
outcome of transfer of embryos derived solely from normal oocytes (group A:
72 patients, 183 embryos) was compared with those from oocytes with
cytoplasmic abnormalities (group B: 34 patients, 85 embryos). In group A,
17 clinical pregnancies (24% per patient, implantation rate 10%) were
established. In group B, only one clinical pregnancy (3% per patient,
implantation rate 1%) was established, from the transfer of embryos derived
from oocytes with homogeneous granularity of the cytoplasm. No pregnancy
resulted following the transfer of embryos from eggs with cytoplasmic
inclusions. The difference was statistically significant. The outcome of
ICSI is dependent on the quality of the oocytes retrieved. Normal
fertilization and early embryo development were achieved in oocytes with
abnormal cytoplasm morphology, but the resulting embryos failed to
demonstrate the same implantation potential as those derived from oocytes
with normal cytoplasm.
相似文献
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Mark J Elder MD FRACS FRACO Paul Hiscott PhD FRCS MRCPath John K.G Dart DM FRCS FRCOphth 《Human pathology》1997,28(12):1348-1354
Cicatricial conjunctivitis may be a sequel to systemic disorders (eg, Stevens-Johnson syndrome, cicatricial pemphigoid) or local disorders such as chemical burns. The cicatrisation is often associated with corneal epithelial changes that cause visual loss. These have been attributed to encroachment of the conjunctival epithelium over the cornea. However, the epithelial anomalies are poorly understood. We investigated the corneal epithelial changes in cicatricial conjunctivitis with an immunohistochemical study of intermediate filaments in normal and pathological specimens. Our results show that the normal corneal epithelium is immunoreactive for cytokeratin 3 (CK 3) but not cytokeratin 19 (CK 19), whereas normal conjunctival epithelium is CK 3 negative and CK 19 positive. Conjunctiva artificially transposed over the cornea (after therapeutic conjunctival flap reconstruction) retained the normal pattern of conjunctival cytokeratin expression (CK 3 negative, CK 19 positive). Conversely, the entire corneal epithelium exhibited the normal cytokeratin pattern (CK 3 positive, CK 19 negative) in 82% of Stevens-Johnson, 80% of cicatricial pemphigoid, and 69% of chemical burns specimens. The findings suggest that conjunctival encroachment is not responsible for the changes at the corneal surface in cicatricial conjunctivitis and that the abnormal corneal epithelium is derived from native corneal cells in these diseases. 相似文献
27.
0 引言 急性乳腺炎是产后妇女的常见病 .我们采用针头挑拨治疗急性乳腺炎 32例 ,疗效显著 .1 临床资料 患者 32例 ,年龄 2 3~ 34 (平均 2 5 .3)岁 ,均为产后哺乳产妇 .其中患乳胀痛为共有症状 .伴发热 10例 ;右侧乳腺炎 14例 ,左侧乳腺炎 16例 ,双侧乳腺炎 2例 ;外上象限胀痛 6例 ,外下象限胀痛 8例 ,内上象限胀痛 7例 ,内下象限胀痛 6例 ,二象限同时胀痛 5例 .乳头均无破损 .患者取卧位 ,显露患乳 ,乳头以碘伏严密消毒后 ,术者持五号针头以指腹常规触诊患乳 ,触摸到肿胀部位后 ,沿乳腺管走行方向找到对应的乳头腺管口 ,以针尖轻轻挑拨… 相似文献
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GA Smith SD Strausbaugh C Harbeck-Weber DM Cohen BJ Shields JD Powers 《Pediatrics》1997,100(5):825-830
OBJECTIVE: To compare the effectiveness of three new topical anesthetics that do not contain cocaine (prilocaine-phenylephrine, tetracaine-phenylephrine [tetraphen], and tetracaine-lidocaine-phenylephrine) to that of tetracaine-adrenaline-cocaine (TAC) during laceration repair in children. DESIGN: Prospective, randomized, double-blind clinical trial. SETTING: The emergency department of an urban children's hospital. PARTICIPANTS: Children 1 year of age or older with a laceration = 5 cm in length that required suturing. Intervention. A total of 240 children were randomly assigned to one of four treatment groups. OUTCOME MEASURES: Pain felt during suturing was scored by suture technicians, research assistants, parents, and patients >/= 5 years of age using a visual analogue scale (VAS). Suture technicians, research assistants, and parents also scored pain using a seven-point Likert scale. In addition, suture technicians completed an anesthetic effectiveness scale. RESULTS: There was consistently no difference demonstrated between the effectiveness of tetraphen and that of TAC for each outcome measure of each observer group. A statistically significant difference was seen among anesthetics when comparing VAS and Likert scale scores of suture technicians and Likert scale scores of research assistants. Based on post hoc analyses, these statistically significant differences were between TAC and prilocaine-phenylephrine (suture technician VAS and Likert scale) and between TAC and tetracaine-lidocaine-phenyl-ephrine (suture technician Likert scale), but not between TAC and tetraphen. When power analyses were performed using alpha = 0.05 and beta = 0.20, it was possible to detect a difference of 1.2 VAS units for each of the observer groups. Based on anesthetic effectiveness scale scores, the three new topical preparations collectively performed significantly better on the face and scalp than on the extremities (relative risk = 1.83; 95% confidence interval 1.20 < relative risk < 2.79). CONCLUSION: This study demonstrated the effectiveness and safety of three new non-cocaine-containing topical anesthetics. Consistently, there was no statistical difference demonstrated between the effectiveness of tetraphen and that of TAC for each outcome measure of each observer group. Tetraphen offers an effective alternative to TAC during laceration repair in children. 相似文献