• Title/Summary/Keyword: Q-FISH

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Combined Study of Cytogenetics and Fluorescence in Situ Hybridization (FISH) Analysis in Childhood Acute Lymphoblastic Leukemia (ALL) in a Tertiary Cancer Centre in South India

  • Mazloumi, Seyed Hashem Mir;Madhumathi, D.S.;Appaji, L.;Prasannakumari, Prasannakumari
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3825-3827
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    • 2012
  • FISH is one of the most sensitive molecular methods to detect genetic abnormalities with DNA probes. When cytogenetic studies are normal or insufficient, FISH may detect cryptic rearrangements, rare or slowly proliferative abnormal populations in non-mitotic cells. We cytogenetically evaluated 70 childhood ALL - 67.1% were found to have an abnormal karyotype. The 23 patients (32.9%) with a normal karyotype were analyzed by FISH applying two probes; TEL/AML1 and MYB which detect cryptic rearrangements of t(12;21)(p13;q22) and deletion of (6q) respectively, associated with a good prognosis. Out of 23 patients, one was positive for t(12;21)(p13;q22) (4.3%). None of our patients were positive for MYB del(6q). Two patients showed an extra signal for MYB on chromosomes other than 6 (8.6 %) indicating amplification or duplication. Findings were compared with the available literature. Our study clearly indicated the integrated FISH screening method to increase the abnormality detection rate in a narrow range. FISH is less useful for diagnostic study of patients with suspected del(6q) but it helps in detecting known cryptic rearrangements as well as identification of new abnormalities(translocation , duplication and amplification) at the gene level.

Prevalence of Haplorchis taichui and Haplorchoides sp. Metacercariae in Freshwater Fish from Water Reservoirs, Chiang Mai, Thailand

  • Nithikathkul, Choosak;Wongsawad, Chalobol
    • Parasites, Hosts and Diseases
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    • v.46 no.2
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    • pp.109-112
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    • 2008
  • A parasitological investigation on trematode metacercariae was made on 62 freshwater fishes of 13 species in northern Thailand; Cyclocheilichthys apogon, Puntioplites proctozysron, Labiobarbus siamensis, Barbodes gonionotus, Barbodes altus, Henicorhynchus siamensis, Osteochilus hasselti, Notopterus notopterus, Mystacoleucus marginatus, Anabas testudineus, Systomus orphoides, Morulius chrysophykadian, and Hampala macrolepidota. The fish were caught over the summer period (February-May 2007) from 2 Chiang Mai water reservoirs, i.e., the Mae Ngad (UTM 47Q E 503200, 47Q N 2119300) and the Mae Kuang Udomtara (UTM 47Q E 513000, 47Q N 2092600) Reservoirs in Chiang Mai province, Thailand. The prevalence of heterophyid (Haplorchis taichui and Haplorchoides sp.) metacercariae in these fish was 83.9% and 74.2% in the Mae Ngad and Mae Kuang Udomtara Reservoirs, respectively. The highest intensity of heterophyid metacercariae in H. siamensis in the Mae Ngad was 120.4 and that in P. proctozysron in the Mae Kuang Udomtara was 180.0. The fish, A. testudineus, C. apogon, and M. chrysophykadian, were not found to be infected with H. taichui metacercariae. The results show that the freshwater fish in Chiang Mai water reservoirs are heavily infected with H. taichui and Haplorchoides sp. metacercariae.

Effects of Dietary Quartz Porphyry and Feed Stimulants, BAISM Supplementation on Growth Performance and Disease Resistance of juvenile eel Anguilla japonica (사료내 맥반석과 BAISM 복합첨가가 치어기 뱀장어 Anguilla japonica의 성장과 내병성에 미치는 영향)

  • Bae, Jun-Young;Han, Kyung-Min;Lee, Jun-Ho;Kim, Sang-Eun;Lee, Jeong-Yeol;Bai, Sung-Chul C.
    • Journal of Aquaculture
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    • v.21 no.1
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    • pp.26-33
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    • 2008
  • This study investigated the synergistic effects of dietary supplementation of quartz porphyry(QP) and a laboratory developed feed stimulants, BAISM(BS) on growth performance and utilization as the additives for juvenile eel Anguilla japonica. Six isoenergetic experimental diets(18.2 kJ/g) were formulated to contain 50% crude protein, 15% lipid with or without dietary QP(Song-Gang stone, Davistone, Korea) and BS supplementation. QP and BS were provided at 0% in the control diet($Q_0B_0$) and at 0.7% QP+0% BS($Q_{0.7}B_0$), 0.7% QP+0.3% BS($Q_{0.7}B_{0.3}$), 0.7% QP+0.5% BS($Q_{0.7}B_{0.5}$), 0.7% QP+0.75% BS($Q_{0.7}B_{0.75}$) and 0.7% QP+1.0% BS($Q_{0.7}B_{1.0}$) in experimental diets on dry matter basis. After four weeks of adaptation, triplicate groups of 30 fish initially averaging $15{\pm}0.1g(mean{\pm}SD)$ were randomly distributed into each aquarium, and they were fed one of the experimental diets for 8 weeks. By the end of the feeding trial, weight gain(%), specific growth rate(%), feed efficiency(%) and protein efficiency ratio of fish fed diet $Q_{0.7}B_{0.5},\;Q_{0.7}B_{0.75}\;and\;Q_{0.7}B_{1.0}$, were significantly higher(P<0.05) than those of fish fed the other diets. But, $Q_{0.7}B_{0.5},\;Q_{0.7}B_{0.75}\;and\;Q_{0.7}B_{1.0}$ were no significant differences(P<0.05). In challenge test, fish were infected by intraperitoneal injection of 0.1 mL bacterial suspension with Edwardsiella tarda per fish after the feeding trial. As a result, fish fed QP and BS supplemented diets have a significantly higher cumulative survival rate than those of fish fed control diet(P<0.05). In conclusion, these results indicated that the optimum dietary supplementation level of QP and BS could be approximately 0.7% quartz porphyry+0.5% BAISM($Q_{0.7}B_{0.5}$) of diet based on WG, FER, SGR, PER, cumulative survival rate in juvenile eel A. japonica.

Detection of HER2 Status in Breast Cancer: Comparison of Current Methods with MLPA and Real-time RT-PCR

  • Pazhoomand, Reza;Keyhan, Elahe;Banan, Mehdi;Najmabad, Hossein;Karimlou, Masoud;Khodadad, Faranak;Iraniparast, Alireza;Feiz, Farnaz;Majidzadeh, Keivan;Bahman, Ideh;Moghadam, Fatemeh Aghakhani;Sobhani, Atoosa Madadkar;Abedin, Seyedeh Sedigheh;Muhammadnejad, Ahad;Behjat, Farkhondeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7621-7628
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    • 2013
  • Human epidermal growth factor receptor (HER) status is an important prognostic factor in breast cancer. There is no globally accepted method for determining its status, and which method is most precise is still a matter of debate. We here analyzed HER2 mRNA expression by quantitative reverse transcription-PCR (qRT-PCR) and HER2 DNA amplification using multiplex ligation-dependent probe amplification (MLPA). In parallel, we performed a routine evaluation of HER2 protein by immunohistochemistry (IHC). To assess the accuracy of the RT-PCR and MLPA techniques, a combination of IHC and fluorescence in situ hybridization (FISH) was used, substituting FISH when the results of IHC were ambiguous (2+) and for those IHC results that disagreed with MLPA and qRT-PCR, this approach being termed IHC-FISH. The IHC results for four samples were not compatible with the MLPA and qRT-PCR results; the MLPA and qRT-PCR results for these samples were confirmed by FISH. The correlations between IHC-FISH and qRT-PCR or MLPA were 0.945 and 0.973, respectively. The ASCO/CAP guideline IHC/FISH correlation with MLPA was (0.827) and with RT-PCR was (0.854). The correlations between the IHC results (0, 1+ as negative, and 3+ as positive) and qRT-PCR and MLPA techniques were 0.743 and 0.831, respectively. Given the shortcomings of IHC analysis and greater correlations between MLPA, qRT-PCR, and FISH methods than IHC analysis alone with each of these three methods, we propose that MLPA and real-time PCR are good alternatives to IHC. However a suitable cut-off point for qRTPCR is a prerequisite for determining the exact status of HER2.

The first Korean case of a newborn with 3p26 microdeletion and 5q35 microduplication inherited from paternal balanced translocation

  • Jang, Jin A;Sohn, Young Bae;Lee, Jang Hoon;Park, Moon Sung
    • Journal of Genetic Medicine
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    • v.18 no.1
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    • pp.48-54
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    • 2021
  • Genetic imbalances are a major cause of congenital and developmental abnormalities. We report the first case of a 3p26 microdeletion and 5q35.2q35.3 microduplication in a newborn with multiple congenital anomalies evaluated using chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH). The patient was born at 30 weeks and 2 days of gestation with a body weight of 890 g. He had symmetric intrauterine growth restriction, microcephaly, facial dysmorphism (hypertelorism, blepharophimosis, mild low-set ears, high-arched palate, and micrognathia), and right thumb polydactyly. Echocardiography revealed an atrial septal defect and patent ductus arteriosus. Furthermore, CMA revealed a concurrent microdeletion in 3p26 and a microduplication in 5q35.2q35.3. FISH analysis showed that these genetic changes resulted from a translocation mutation between chromosomes 3 and 5. The patient's mother had mild intellectual disability, short stature, and facial dysmorphism, while his father had a normal phenotype. However, parental FISH analysis revealed that the asymptomatic father carried a balanced translocation of chromosomes 3p26 and 5q35. CMA and FISH tests are useful for diagnosing neonates with multiple congenital abnormalities. Further parental genetic investigation and proper genetic counseling are necessary in cases of chromosomal abnormalities inherited from parental balanced translocations.

Importance of FISH combined with Morphology, Immunophenotype and Cytogenetic Analysis of Childhood/Adult Acute Lymphoblastic Leukemia in Omani Patients

  • Goud, Tadakal Mallana;Al Salmani, Kamla Khalfan;Al Harasi, Salma Mohammed;Al Musalhi, Muhanna;Wasifuddin, Shah Mohammed;Rajab, Anna
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7343-7350
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    • 2015
  • Genetic changes associated with acute lymphoblastic leukemia (ALL) provide very important diagnostic and prognostic information with a direct impact on patient management. Detection of chromosome abnormalities by conventional cytogenetics combined with fluorescence in situ hybridization (FISH) play a very significant role in assessing risk stratification. Identification of specific chromosome abnormalities has led to the recognition of genetic subgroups based on reciprocal translocations, deletions and modal number in B or T-cell ALL. In the last twelve years 102 newly diagnosed childhood/adult ALL bone marrow samples were analysed for chromosomal abnormalities with conventional G-banding, and FISH (selected cases) using specific probes in our hospital. G-banded karyotype analysis found clonal numerical and/or structural chromosomal aberrations in 74.2% of cases. Patients with pseudodiploidy represented the most frequent group (38.7%) followed by high hyperdiploidy group (12.9%), low hyperdiploidy group (9.7%), hypodiploidy (<46) group (9.7%) and high hypertriploidy group (3.2%). The highest observed numerical chromosomal alteration was high hyperdiploidy (12.9%) with abnormal karyotypes while abnormal 12p (7.5%) was the highest observed structural abnormality followed by t(12;21)(p13.3;q22) resulting in ETV6/RUNX1 fusion (5.4%) and t(9;22)(q34.1;q11.2) resulting in BCR/ABL1 fusion (4.3%). Interestingly, we identified 16 cases with rare and complex structural aberrations. Application of the FISH technique produced major improvements in the sensitivity and accuracy of cytogenetic analysis with ALL patients. In conclusion it confirmed heterogeneity of ALL by identifying various recurrent chromosomal aberrations along with non-specific rearrangements and their association with specific immunophenotypes. This study pool is representative of paediatric/adult ALL patients in Oman.

Molecular cloning and foreign gene expression of restriction endonuclease fragments of the Hc nuclear polyhedrosis virus DNA (Hc nuclear polyhedrosis virus DNA 제한효소절편의 molecular cloning 과 외래 유전자 발현)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.8 no.1
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    • pp.31-36
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    • 1995
  • Hc nuclear polyhedrosis virus DNA genome was digested with EcoRI endonuclease, these partial fragments were recombined into the pUC8 plasmid vector and transformed in E. coli JM 83 cell. The genome DNA has 24 EcoRI fragments and 12 fragments of them were subcloned. The four recombinants were named as eNP-O, eNP-Q, eNP-R and eNP-S. The expression of foregin gene of the recombinants was investigated by analysing protein patterns on the SDS-PAGE. The eNP-O, eNP-Q and eNP-R were expressed a different weight of protein as comparision with potypeptide bands of E. coli JM 83 host cell.

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Comparative Analysis of Market Demand and Individual Demand for Major Fish Species in Korea (한국 주요 어종의 시장수요와 개인수요의 비교분석)

  • Park, Hoan-Jae
    • The Journal of Fisheries Business Administration
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    • v.43 no.1
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    • pp.35-48
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    • 2012
  • Inverse demand models are well established as market demands in theory and practice of the existing literature. However, the derivation and its interpretation of individual demands from the market demands are not well known in the literature. This paper analyzes the fish market in Korea by the inverse demand model and shows how we deduce the consumer's responses from the market responses when the markets determine the prices by the quantities demanded. It illustrates empirically how this can be done applying to the korean fish market data. The empirical results show that all fishes are price inflexible and mackerels and hairtails are scale flexible in the market demand while mackerels, hairtails, and croakers are price elastic and mackerels and hairtails are income inelastic in the individual demand. The methodology and empirics used in the paper will make a contribution to the existing literature especially for the purpose of recovering consumer's demand from the market demand, thus implementing the policies to administer the fish markets.

Chromosomal Localization and Distribution of the Telomeric DNA in Cattle and Pigs (소, 돼지 염색체의 telomeric DNA 분포 양상)

  • Sohn, S.H.;Multani, A.S.;Pathak, S.;Cho, E.J.;Ha, H.B.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.547-554
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    • 2004
  • Telomeres are nucleoprotein structures at the ends of chromosomes consisting of tandem repeat sequences of . (TTAGGG)n. Telomeres serve as guardians of the genome, protect individual chromosomes within the nucleus, and help in meiotic pairing of homologous chromosomes. To investigate the telomere distributions of cattle and pig chromosomes, fluorescence in situ hybridization(FISH) was carried out on metaphase spreads of in vitro fibroblast cultures from Holstein and Landrace using a human telomeric DNA repeat probe. Results indicate that the distinct double spots on both ends of chromosomes of cattle and pigs were observed. In cattle, there was a random variation in the intensity of telomere signals among chromosomes. In pigs, an interstitial telomeric signal was observed on the chromosome 6q1 of all the cells examined. According to quantitative fluorescence in situ hybridization(Q-FISH) analysis, some chromosomes had consistently much more telorneres at one end of chromosomes. In general, both species had consistently much more telomeres at q-end than p-end on most of chromosomes. The relative amount of telomeres on bovine chromosomes was higher than that on pig chromosomes. In additions, Y chromosome had the highest relative amount of telorneres in cattle and pigs.

14q32.33 Deletion Identified by array-CGH in a 5-year old-girl with Seizure

  • Cheon, Chong-Kun;Park, Sang-Jin;Choi, Ook-Hwan
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.62-66
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    • 2011
  • Deletions of 14q including band 14q32.33 are uncommon. Patients with terminal deletions of chromosome 14 usually share a number of clinical features. By molecular techniques (array comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH), we identified a young girl with 0.3 Mb terminal 14q32.33 deletion. Review of the nine cases with pure terminal 14q32.3 deletions described to date documented that our observation is the smallest terminal 14q deletion ever reported. The phenotype of our patient is much less severe than the phenotypes of the patients reported previously. We report our experience in examining the clinical, behavioral, and cognitive findings in a 5-year-old girl studied with chromosomal microarray hybridization and reviewed previously reported patients with 14q32 deletions.