scholarly journals DETEKSI MOLEKULER PENYEBAB PENYAKIT KUNING (Tomato chlorosis virus DAN Tomato infectious chlorosis virus) PADA TANAMAN TOMAT (MOLECULAR DETECTION CAUSED YELLOWING DISEASE (Tomato chlorosis virus AND Tomato infectious chlorosis virus) ON TOMATOES)

2017 ◽  
Vol 19 (2) ◽  
pp. 80
Author(s):  
Resti Fajarfika ◽  
Sedyo Hartono ◽  
Sri Sulandari ◽  
Susamto Somowiyarjo
Keyword(s):  
Molecular Detection ◽  
Nucleotide Sequence Analysis ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Pcr Product ◽  
Yellowing Disease ◽  
Polymerase Chain ◽  
Tomato Chlorosis Virus ◽  
Tomato Infectious Chlorosis Virus

ABSTRACTThis research was aimed to detect the ToCV and TICV caused yellowing disease on tomatoes by molecular detection. Leaf samples of symptomatic plants were taken from Ketep (Magelang), then the leaves were identified by reversetranscription-polymerase chain reactions (RT-PCR) using specific primer ToCV-CF/ToCV-CR (360 bp) and TICVCF/TICV-CR (416 bp). The result of nucleotide sequence analysis, amino acid and PCR product phylogenetic sequences were verified as TICV, it showed that TICV from Magelang belongs to the same group with TICV from Japan, North America and Europe, France, Italy, and USA.Keywords: molecular detection, ToCV, TICVINTISARIPenelitian ini bertujuan untuk mendeteksi keberadaan ToCV dan TICV penyebab penyakit kuning pada tanaman tomat. Daun bergejala diambil dari Desa Ketep (Magelang), selanjutnya diuji denganreverse transcription-polymerasechain reactions(RT-PCR) menggunakan primer spesifik ToCV-CF/ToCV-CR (360 bp) dan TICV-CF/TICV-CR (416 bp). Hasil analisis sekuen nukleotida, asam amino, dan filogenetik produk PCR teridentifikasi sebagai TICV yang menunjukkan bahwa TICV isolat Magelang berada dalam satu kelompok dengan isolat TICV asal Jepang, Amerika Utara dan Eropa,Perancis, Italia, dan USA.Kata kunci: deteksi molekuler, ToCV, TICV

scholarly journals Characteristic Differences in Reverse Transcription-Polymerase Chain Reaction Products of Ovine, Bovine, and Human Respiratory Syncytial Viruses

1993 ◽  
Vol 5 (3) ◽  
pp. 322-328 ◽  
Author(s):  
Richard D. Oberst ◽  
Michael P. Hays ◽  
Jim F. Evermann ◽  
Clayton L. Kelling
Keyword(s):  
Reverse Transcription ◽  
Reaction Products ◽  
Rt Pcr ◽  
Specific Primers ◽  
F Protein ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Respiratory Syncytial Viruses ◽  
Polymerase Chain ◽  
Syncytial Virus

In reverse transcription-polymerase chain reactions (RT-PCR) and DNA hybridizations using primers and an oligonucleotide probe to the fusion (F) protein mRNA of bovine respiratory syncytial virus (BRSV), all the BRSV isolates and a goat isolate could be distinguished from prototype isolates of human respiratory syncytial viruses (HRSV) and ovine (sheep and bighorn sheep) respiratory syncytial viruses (RSV). However, RT-PCR amplifications with primers to sequences of the HRSV F protein mRNA resulted in amplified products of ≍ 243 bp if mRNA templates of subgroup A HRSV strains were present and slightly larger amplified products with subgroup B HRSV strains. No amplified products were observed in HRSV-primed RT-PCR with BRSV or goat or ovine RSV mRNA templates. Although the ovine RSV isolates were antigenically cross-reactive with the goat RSV, HRSV and BRSV isolates, they were not amplified with either HRSV- or BRSV-specific primers in RT-PCR. These results confirm previous immunological comparisons suggesting that some ovine RSV isolates should be considered as distinct respiratory syncytial viruses.

Detection of rye chromosome 2R using the polymerase chain reaction and sequence-specific DNA primers

Genome ◽  
1994 ◽  
Vol 37 (1) ◽  
pp. 19-22 ◽  
Author(s):  
J.-H. Lee ◽  
R. A. Graybosch ◽  
D. J. Lee
Keyword(s):  
Polymerase Chain Reaction ◽  
Wheat Chromosome ◽  
Specific Marker ◽  
Chain Reaction ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Wheat Lines ◽  
Dna Primers

Sequences derived from a rye gamma secalin gene were used as primers in polymerase chain reactions using DNA obtained from a series of wheat and triticale genetic stocks. A 473-bp fragment, the predicted size based on the distance between the selected primers, was found only in rye, triticales, and wheat lines carrying rye chromosome 2RS. Use of triticale lines with various wheat chromosome substitutions confirmed the chromosomal origin of the rye-specific marker. The presence of the 473-bp PCR product was always associated with the production of 75K secalins in grain samples. Thus, the primer sequences, and the clone of origin (pSC503), were both derived from the SEC-2 locus of rye chromosome 2RS.Key words: wheat (Triticum aestivum), rye (Secale cereale), chromosomal translocations, chromosomal substitutions, DNA polymerase chain reaction, sequence-specific primers.

scholarly journals SARS-CoV-2 show no infectivity at later stages in a prolonged COVID-19 patient despite positivity in RNA testing

2021 ◽  
Author(s):  
Xiu-Feng Wan ◽  
Cynthia Y Tang ◽  
Detlef Ritter ◽  
Yang Wang ◽  
Tao Li ◽  
...  
Keyword(s):  
Large Scale ◽  
Disease Onset ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Virus Growth ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Asymptomatic Patients ◽  
Hospitalization Period ◽  
Polymerase Chain

Inpatient COVID-19 cases present enormous costs to patients and health systems. Many hospitalized patients may still test COVID-19 positive, even after resolution of symptoms. Thus, a pressing concern for clinicians is the safety of discharging these asymptomatic patients if they have any remaining infectivity. This case report explores the viral viability in a patient with persistent COVID-19 over the course of a two-month hospitalization. Positive nasopharyngeal swab samples, analyzed by quantitative reverse transcription polymerase chain reactions (qRT-PCR), were collected and isolated in the laboratory, and infectious doses were analyzed throughout the hospitalization period. The patient experienced waning symptoms by hospital day 40 and had no viable virus growth in the laboratory by hospital day 41, suggesting no risk of infectivity, despite positive RT-PCR results, which prolonged his hospital stay. Notably, this case showed infectivity for at least 24 days from disease onset, which is longer than the discontinuation of transmission-based precautions recommendation by CDC. Thus, our findings suggest that the timeline for discontinuing transmission-based precautions may need to be extended for patients with prolonged illness. Additional large-scale studies are needed to draw definitive conclusions on the appropriate clinical management for these patients.

scholarly journals Prognostic value of Serum miR-217 Level in Osteosarcoma Patients

2020 ◽  
Author(s):  
Jia Ye ◽  
Zhi-Hui Jin ◽  
Ren Chen ◽  
Sen Chen ◽  
Yi-Jun Ren ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Poor Prognosis ◽  
Prognostic Significance ◽  
Serum Levels ◽  
High Serum ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Low Serum

Abstract Background MicroRNA (miR)-217 is a tumor suppressor significantly associated with osteosarcoma. We try to evaluate serum levels miR-217 in osteosarcoma patients and evaluate its prognostic significance. Methods A total of 163 consecutive osteosarcoma patients and 96 healthy participates were enrolled. Serum miR-217 levels were evaluated by using real-time quantitative reverse transcription polymerase chain reactions(RT-PCR). The association between serum miR-217 level and survival outcomes was evaluated by univariate and multivariate analysis. Results Serum miR-217 levels in osteosarcoma patients was significantly lower than healthy volunteers (P<0.05). Low serum miR-217 was significantly related to advanced cancer and metastasis (both P<0.05). Moreover, patients with a low serum miR-217 had a poorer overall survival than those with a high serum miR-217 levels (P<0.05). Serum miR-217 level also been showed as independent risk factor for osteosarcoma in multivariate analysis (HR, 0.42; 95%CI: 0.12–0.98; p<0.01). Conclusions Serum miR-217 levels was significantly downregulated in osteosarcoma patients and remarkably associated with poor prognosis, indicating that serum miR-217 might serve as a useful diagnostic and prognostic indictor for osteosarcoma.

Detection of phytoplasmas in dieback, yellow crinkle, and mosaic diseases of papaya using polymerase chain reaction techniques

1996 ◽  
Vol 47 (3) ◽  
pp. 387 ◽  
Author(s):  
B Liu ◽  
DT White ◽  
KB Walsh ◽  
PT Scott
Keyword(s):  
Rrna Genes ◽  
23S Rrna ◽  
Rrna Gene ◽  
Tissue Samples ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Pcr Product ◽  
Polymerase Chain ◽  
23S Rrna Genes ◽  
The 16S Rrna Gene

Oligonucleotide primers complementary to regions specific to plant-pathogenic mycoplasma-like organisms (phytoplasmas) were used in polymerase chain reactions on tissue samples from dieback, yellow crinkle, and mosaic affected papaya plants. The primer pair P068/P069, which hybridise to internal regions of the 16s rRNA gene, amplified an approximately 560 bp product in dieback, yellow crinkle and mosaic affected papaya. The primer pair P3/P7, which hybridise to the spacer region between the 16s and 23s rRNA genes, amplified an approximately 300 bp fragment in yellow crinkle and mosaic affected papaya, with no product from dieback affected plants. No PCR product was obtained with either set of primers from healthy plants. An identical Alu I restriction enzyme profile was obtained with all three 560 bp products. This study provides the first evidence for the association of phytoplasmas with papaya mosaic and Australian papaya dieback.

scholarly journals First Report of Tomato infectious chlorosis virus in Tomato in Indonesia

Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 872-872 ◽  
Author(s):  
J. Th. J. Verhoeven ◽  
T. M. Willemen ◽  
J. W. Roenhorst ◽  
R. A. A. van der Vlugt
Keyword(s):  
The United States ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Seedlings ◽  
Large Numbers ◽  
Lateral Shoots ◽  
Young Leaves ◽  
Polymerase Chain ◽  
Tomato Chlorosis Virus ◽  
Tomato Infectious Chlorosis Virus

In 2002, a breeding company submitted several samples of tomato (Lycopersicon esculentum) for diagnosis. Samples originated in Indonesia and were taken from protected and nonprotected crops. Plants exhibited severe chlorosis on fully expanded leaves, while young leaves were symptomless. Symptoms resembled those of the criniviruses Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV). Moreover, large numbers of whiteflies, potential vectors of these viruses, had been observed at the plots with symptomatic plants. A reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for TICV (1) yielded amplicons of the expected size of approximately 500 bp for all samples. One of the amplicons was sequenced (Genbank Accession No. AY221097) and revealed more than 98.9% identity to six isolates of TICV in NCBI Genbank. cDNA synthesis using the universal crinivirus primer HSP_M2-DW (5′ -TCRAARGTWCCKCCNCCRAA-3′) followed by PCR with a ToCV specific primerset (ToCV-UP 5′-TCATTAAAACTCAATGGGACCGAG-3′ and ToCV-DW 5′-GCGACGT AAATTGAAACCC-3′) was negative in all cases. Grafting of symptomatic shoots onto healthy tomato seedlings of cv. Money-maker showed transmission of the virus, as chlorosis appeared on fully expanded leaves of lateral shoots after 6 weeks. The presence of TICV in the graft-inoculated plants was confirmed by RT-PCR. Furthermore, mechanical inoculation to a range of herbaceous test plants did not evoke any virus symptoms, indicating the absence of mechanically transmissible viruses. Although other nonmechanically transmissible viruses cannot be fully excluded, the results together with the symptoms observed, indicate that TICV is the cause of the disease. TICV has been reported from Greece, Italy, Japan, Spain, and the United States, but to our knowledge, this is the first report of TICV in Indonesia. Reference: (1) A. M. Vaira et al. Phytoparasitica 30:290, 2002.

scholarly journals Prognostic value of Serum miR-217 Level in Osteosarcoma Patients

2021 ◽  
Author(s):  
Jia Ye ◽  
Zhi-Hui Jin ◽  
Ren Chen ◽  
Sen Chen ◽  
Yi-Jun Ren ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Prognostic Significance ◽  
Serum Levels ◽  
Prognostic Indicator ◽  
High Serum ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Low Serum

Abstract Background: MicroRNA (miR)-217 is a tumor suppressor significantly associated with osteosarcoma. We try to evaluate serum levels miR-217 in osteosarcoma patients and evaluate its prognostic significance.Methods: A total of 163 consecutive osteosarcoma patients and 96 healthy participates were enrolled. Serum miR-217 levels were evaluated by using real-time quantitative reverse transcription polymerase chain reactions(RT-PCR). The association between serum miR-217 level and survival outcomes was evaluated by univariate and multivariate analysis. Results: Serum miR-217 levels in osteosarcoma patients was significantly lower than healthy volunteers (P<0.05). Low serum miR-217 was significantly related to advanced cancer and metastasis (both P<0.05). Moreover, patients with a low serum miR-217 had a poorer overall survival than those with a high serum miR-217 levels (P<0.05). Serum miR-217 level also been showed as independent risk factor for osteosarcoma in multivariate analysis (HR, 0.42; 95%CI: 0.12–0.98; p<0.01). Conclusions: Serum miR-217 levels was significantly downregulated in osteosarcoma patients and remarkably associated with poor prognosis, indicating that serum miR-217 might serve as a useful diagnostic and prognostic indicator for osteosarcoma.

Identification of the NO Synthase isoforms Expressed in Human Neutrophil Granulocytes, Megakaryocytes and Platelets

1997 ◽  
Vol 77 (01) ◽  
pp. 163-167 ◽  
Author(s):  
Thomas Wallerath ◽  
Ingolf Gath ◽  
Walter E Aulitzky ◽  
Jennifer S Pollock ◽  
Hartmut Kleinert ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Platelets ◽  
No Synthase ◽  
Immunofluorescent Staining ◽  
Human Neutrophils ◽  
Neutrophil Granulocytes ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain

SummaryUsing Western blot and fluorescent immunocytochemistry, NOS III (or ecNOS) and NOS II (or iNOS), but no NOS I (or ncNOS), were identified in preparations of human platelets. Reverse-transcription polymerase chain reactions (RT-PCR) demonstrated NOS III mRNA, but no NOS II mRNA (which is short-lived) and no NOS I mRNA in platelets. Immunofluorescent staining of human bone marrow smears showed the presence of NOS III, but not NOS I in megakaryocytes. A subpopulation of megakaryocytes also expressed NOS II. In preparations of human neutrophils, immunocytochemistry demonstrated NOS I in all cells, whereas no NOS III was detected. The few NOS II positive cells were characterized as contaminating eosinophils. Similarly, in RT-PCR, transcripts for NOS I and NOS II, but not for NOS III, were identified. Thus, the constitutive NOS isoform in megakaryocytes and platelets is NOS III, whereas neutrophils express NOS I. Some megakaryocytes and eosinophils also express NOS II.

scholarly journals Management of Severe COVID-19 in Pregnancy

2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Yassamine Abourida ◽  
Houssam Rebahi ◽  
Imane Oussayeh ◽  
Hajar Chichou ◽  
Bouchra Fakhir ◽  
...  
Keyword(s):  
Severe Pneumonia ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Premature Rupture Of Membranes ◽  
Premature Rupture ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Pregnancy And Delivery ◽  
Polymerase Chain ◽  
Core Problem

The scarcity of data concerning pregnant patients gravely infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) makes their management difficult, as most of the reported cases in the literature present mild pneumonia symptoms. The core problem is laying out evidence on coronavirus’s implications on pregnancy and delivery, as well as vertical transmission and neonatal mortality. A healthy 30-year-old pregnant woman, gravida 6, para 4, at 31 weeks of gestation, presented severe pneumonia symptoms promptly complicated with premature rupture of membranes (PROM). A nasopharyngeal swab returned positive for SARS-CoV-2 using reverse transcription polymerase chain reactions (RT-PCR). The parturient underwent a cesarean delivery. This paper is an attempt to outline management of the critical condition of COVID-19 during pregnancy.

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