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Nancy B Kiviat - One of the best experts on this subject based on the ideXlab platform.
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comparison of dot Filter Hybridization southern transfer Hybridization and polymerase chain reaction amplification for diagnosis of anal human papillomavirus infection
Journal of Clinical Microbiology, 1993Co-Authors: Jane Kuypers, Cathy W Critchlow, Debra Vernon, P E Gravitt, James Sayer, M M Manos, Nancy B KiviatAbstract:The detection and classification of human papillomavirus (HPV) by a consensus primer polymerase chain reaction (PCR) technique were compared with detection and classification by dot Filter Hybridization (DFH) and Southern transfer Hybridization (STH). PCR detected HPV in 87% of specimens; the detection rates for DFH and STH were 51% and 49%, respectively. The specific HPV types detected by STH were also detected by PCR in 90% of specimens. However, 75% of the samples positive for unclassified HPV by STH were typed by PCR. PCR results were reproducible, as assessed by repeat analysis (96% agreement), by analysis of paired same-day specimens (89% agreement), and by interlaboratory analysis (88% agreement). PCR is a sensitive, specific, and reproducible test for HPV detection and classification in clinical and epidemiologic studies.
Olli Ritvos - One of the best experts on this subject based on the ideXlab platform.
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regulation of inhibin alpha and beta a subunit messenger ribonucleic acid levels by chorionic gonadotropin and recombinant follicle stimulating hormone in cultured human granulosa luteal cells
The Journal of Clinical Endocrinology and Metabolism, 1994Co-Authors: Marja Eramaa, Timo Tuuri, Kristiina Hilden, Olli RitvosAbstract:We studied the effects of recombinant human FSH (rhFSH) and purified hCG on the steady state messenger ribonucleic acid (mRNA) levels of inhibin alpha- and beta A-subunits in cultured granulosa-luteal cells of preovulatory ovarian follicles obtained from women undergoing in vitro fertilization. Specific mRNA transcripts for the alpha- and beta A-subunits were detected in Northern and dot blot Filter Hybridization analyses, and the levels of these mRNAs were induced by rhFSH and hCG in a distinct concentration- and time-dependent manner. The basal and hCG-stimulated alpha-subunit mRNA levels were first determined at 2- to 3-day intervals over a 3- to 10-day culture period after the initiation of the cultures. Both the basal and hCG-stimulated alpha-subunit mRNA levels declined steadily during culture, but the maximal relative stimulatory effect of hCG was observed on day 7 of culture. All subsequent experiments, therefore, were performed on days 6-8 of culture. Both gonadotropins induced alpha-subunit mRNA...
Alan Mclachlan - One of the best experts on this subject based on the ideXlab platform.
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RNA (Northern) Filter Hybridization and RT-qPCR analysis of HBV transcripts in the livers of adult HBV transgenic mice.
2017Co-Authors: Vanessa C. Mcfadden, Rasha E. Shalaby, Saira Iram, Claudia E. Oropeza, Jennifer A. Landolfi, Alexander V. Lyubimov, Mark Maienschein-cline, Stefan J. Green, Klaus H. Kaestner, Alan MclachlanAbstract:(A) RNA (Northern) Filter Hybridization analysis of representative mice of each sex and genotype are shown. Noncontiguous lanes from multiple analysis are presented. The probes used were HBVayw genomic DNA plus GAPDH cDNA. FoxA-expressing (HBVFoxA2fl/flAlbCre(-), HBVFoxA1fl/flFoxA2fl/flAlbCre(-) and HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(-)) and FoxA-deleted (HBVFoxA2fl/flAlbCre(+), HBVFoxA1fl/flFoxA2fl/flAlbCre(+) and HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(+)) HBV transgenic mice are indicated (Genotype A2, A1A2 and A1A2A3, respectively). The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) transcript was used as an internal control for the quantitation of the HBV 3.5kb RNA. (B) Quantitative analysis by RNA (Northern) Filter Hybridization of the HBV 3.5kb transcript in the HBV transgenic mice. The mean HBV 3.5kb transcript levels plus standard deviations are indicated. Average number of mice per group was 6.6±1.9 (Range: 4–9). The levels of the HBV 3.5kb transcript which are statistically significantly different between Cre(-) and Cre(+) HBV transgenic mice by a Student’s t-test (p
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RNA (Northern) Filter Hybridization and RT-qPCR analysis of HBV transcripts in the livers of 1, 2, 3, 4 and 7 week old HBV transgenic mice.
2017Co-Authors: Vanessa C. Mcfadden, Rasha E. Shalaby, Saira Iram, Claudia E. Oropeza, Jennifer A. Landolfi, Alexander V. Lyubimov, Mark Maienschein-cline, Stefan J. Green, Klaus H. Kaestner, Alan MclachlanAbstract:(A) RNA (Northern) Filter Hybridization analysis of representative mice of each genotype are shown. Noncontiguous lanes from multiple analysis are presented. The probes used were HBVayw genomic DNA plus GAPDH cDNA. FoxA-expressing (HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(-)) and FoxA-deleted (HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(+)) HBV transgenic mice are indicated (Genotype A1A2A3). The glyceraldehyde 3-phosphate dehydrogenase (GAPDH) transcript was used as an internal control for the quantitation of the HBV 3.5kb RNA. (B) Quantitative analysis by RNA (Northern) Filter Hybridization of the HBV 3.5kb transcript in the HBV transgenic mice. The mean HBV 3.5kb transcript levels plus standard deviations are indicated. The levels of the transcripts which are statistically significantly different between Cre(-) and Cre(+) HBV transgenic mice by a Student’s t-test (p
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DNA (Southern) Filter Hybridization analysis of HBV DNA replication intermediates in the livers of adult HBV transgenic mice.
2017Co-Authors: Vanessa C. Mcfadden, Rasha E. Shalaby, Saira Iram, Claudia E. Oropeza, Jennifer A. Landolfi, Alexander V. Lyubimov, Mark Maienschein-cline, Stefan J. Green, Klaus H. Kaestner, Alan MclachlanAbstract:(A) DNA (Southern) Filter Hybridization analysis of representative mice of each sex and genotype are shown. Noncontiguous lanes from multiple analysis are presented. The probe used was HBVayw genomic DNA. FoxA-expressing (HBVFoxA2fl/flAlbCre(-), HBVFoxA1fl/flFoxA2fl/flAlbCre(-) and HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(-)) and FoxA-deleted (HBVFoxA2fl/flAlbCre(+), HBVFoxA1fl/flFoxA2fl/flAlbCre(+) and HBVFoxA1fl/flFoxA2fl/flFoxA3+/-AlbCre(+)) HBV transgenic mice are indicated (Genotype A2, A1A2 and A1A2A3, respectively). The HBV transgene (Tg) was used as an internal control for the quantitation of the HBV replication intermediates. Tg = HBV transgene; RC = HBV relaxed circular replication intermediates; SS = HBV single stranded replication intermediates. (B) Quantitative analysis of the HBV DNA replication intermediate (RI) levels in HBV transgenic mice. The mean DNA replication intermediate levels plus standard deviations are indicated. Average number of mice per group was 6.7±1.8 (Range: 4–9). The levels of replication intermediates which are statistically significantly different between Cre(-) and Cre(+) HBV transgenic mice by a Student’s t-test (p
Jacques Seydoux - One of the best experts on this subject based on the ideXlab platform.
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the comparative test performance of dot Filter Hybridization viratype and conventional morphologic analysis to detect human papillomavirus
American Journal of Clinical Pathology, 1992Co-Authors: Jonathan Weintraub, Mireille Redard, Jacques SeydouxAbstract:To investigate the test performance of a commercially available detection kit for human papillomavirus (HPV), the relationship between the detection of HPV by dot Filter Hybridization (DFH) and by standard morphologic methods was studied. Four hundred two cervical samples taken from 381 patients referred to a colposcopy clinic were examined. Human papillomavirus DNA sequences were identified and typed using commercially available anti-sense RNA probes. Simultaneous cytologic smears were obtained in 289 patients, directed biopsy samples in 284, and both smears and biopsy samples in 171 samples. Human papillomavirus DNA was detected in 164 specimens (41%), of which 24 (15%) were type 6/11, 74 (45%) were type 16/18, 39 (24%) were type 31/33/35, and 27 (16%) were untyped due to the presence of multiple positive signals. Viral types 16/18 and 31/33/35 were eight and six times more frequent in cervical intraepithelial neoplasia (CIN) II/CIN III lesions than in condyloma/CIN I, respectively. When the cytologic diagnosis was considered the standard of reference, the results of DFH for the detection of HPV were concordant in 167 (56%) paired samples. The sensitivity of DFH was 48% and the specificity was 77%. The distribution of the morphologic diagnoses in the group of false-negative results and true-positive results was similar. When the histologic diagnosis was considered the standard of reference, the efficiency of DFH was 62%, the sensitivity was 59%, and the specificity was 79%. In the subgroup of 118 samples with simultaneous smear and biopsy and at least one positive examination, 42 (36%) were positive by all three methods, 42 (36%) by two, and 34 (29%) by one, including 6 (5%) by DFH alone. Fifteen cases more were detected by the complementary use of DFH and cytology than with cytology alone. The results demonstrated that the sets of patients positive for HPV when detected by DFH or by morphologic methods were not identical but rather overlapped. The detection of HPV may be slightly improved by using DFH in addition to conventional examinations. A significant number of HPV-positive patients without a morphologic lesion and patients with low-grade lesions had HPV 16/18 or 31/33/35, suggesting a possible role for typing in establishing a risk profile. However, given uncertainties in understanding the biology of HPV-associated lesions, the role, if any, of clinical testing for HPV by DFH remains to be defined.
Jane Kuypers - One of the best experts on this subject based on the ideXlab platform.
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comparison of dot Filter Hybridization southern transfer Hybridization and polymerase chain reaction amplification for diagnosis of anal human papillomavirus infection
Journal of Clinical Microbiology, 1993Co-Authors: Jane Kuypers, Cathy W Critchlow, Debra Vernon, P E Gravitt, James Sayer, M M Manos, Nancy B KiviatAbstract:The detection and classification of human papillomavirus (HPV) by a consensus primer polymerase chain reaction (PCR) technique were compared with detection and classification by dot Filter Hybridization (DFH) and Southern transfer Hybridization (STH). PCR detected HPV in 87% of specimens; the detection rates for DFH and STH were 51% and 49%, respectively. The specific HPV types detected by STH were also detected by PCR in 90% of specimens. However, 75% of the samples positive for unclassified HPV by STH were typed by PCR. PCR results were reproducible, as assessed by repeat analysis (96% agreement), by analysis of paired same-day specimens (89% agreement), and by interlaboratory analysis (88% agreement). PCR is a sensitive, specific, and reproducible test for HPV detection and classification in clinical and epidemiologic studies.