Quantitative detection of human immunodeficiency virus type 1 (HIV-1) viral load by SYBR green real-time RT-PCR technique in HIV-1 seropositive patients

Gibellini, Davide; Vitone, Francesca; Gori, Emanuele; Plaça, M. La; Re, Maria Carla

doi:10.1016/j.jviromet.2003.09.030

Cited by 67 publications

(38 citation statements)

References 27 publications

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“…Reverse transcription was per formed using the SYBR PrimeScript RT reagent kit accor ding to the manufacturer's instructions (TaKaRa Bio technolo gy Co., Ltd, Dalian, China). The reaction mixture (10 mL) included 2 mL of 5×PrimeScript Buffer, 0.5 mL of Prime Script RT Enzyme MixI, 0.5 mL of Oligo dT Primer (50 mm/L), 0.5 mL of Random 6 mers (100 mm/L), 6.50 mL of total RNA, and the RT reaction conditions were as follows: 37 °C for 15 min and 85 °C for 5 s. Quantitative real-time PCR was carried out using the fluorescent dye SYBR Green I according to the procedures previously described (10,11). The primers were designed with the primer design software (PerkinElmer Life Sciences, Waltham, MA, USA) and are shown in Table I.…”

Section: Real-time Quantitative Rt-pcrmentioning

confidence: 99%

TNF-α, HGF and TGF-β1 are Involved in Liver Regeneration Following Partial Hepatectomy Using Portal Vein Arterializations

Niu¹,

Dong²,

Zhang³

et al. 2012

Journal of Medical Biochemistry

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Kratak sadr`aj: Eksperimenti vezani za regeneraciju jetre posle parcijalne hepatektomije pokazali su da TNF-a, HGF i TGF-b1 kao i drugi citokini igraju va`ne uloge u razli~itim faza ma regeneracije jetre, me|utim, uticaj arterijalizacije port ne vene (APV) na ekspresiju ovih citokina tokom regene racije jetre nije jasan. Sprague Dawley pacovi nasumi~no su podeljeni u APV grupu i kontrolne grupe, a krv za detekciju ALT uzeta je pomo}u automatskog biohemijskog analiza tora. Ekspresije TNF-a, HGF i TGF-b1 u tkivima jetre detektovane su kvantitativnom RT-PCR. Nivoi ALT u obe grupe u po~etnom periodu posle operacije bili su zna~ajno vi{i nego pre hirur{kog za hvata i postepeno su se normalizovali posle 7 dana od operacije. Posle 12 h i 24 h od zahvata, ekspresija TNF-a u APV grupi bila je zna~ajno vi{a nego u kontrolnoj grupi (P<0,05), ali posle 7 dana od operacije nije uo~ena zna~ajna razlika izme|u dve grupe. Ekspresija HGF u APV grupi posle 12 h bila je sli~na onoj u kontrolnoj grupi, ali je posle 24 h bila zna~ajno vi{a nego u kontrolnoj grupi (P<0,05). Posle 24 h, ekspresija TGF-b1 u APV grupi bila je zna~ajno ni`a nego u kontrolnoj grupi (P<0,05), ali posle 48 h od operacije izme|u dve grupe nije otkrivena zna~ajna razlika. Pozitivan uticaj na arterija li zaciju portne vene u ranim fazama regeneracije jetre bio je povezan s promenama u ekspresijama TNF-a, HGF i TGF-b1. Summary: Experiments on liver regeneration after partial hepatectomy have shown that TNF-a, HGF and TGF-b1 and other cytokines play important roles in the different stages of liver regeneration, however, the effect of portal vein arteria liz ation (PVA) on the expressions of these cytokines during liver regeneration is not clear. Sprague Dawley rats were randomly divided into the PVA group and control groups, and blood was collected for the detection of ALT using an automatic bio chemical analyzer. The expressions of TNF-a, HGF and TGF-b1 in liver tissues were detected by quan titative RT-PCR. The ALT levels in both groups in the early period after surgery were significantly higher than those before operation, and gradually returned to normal at 7 days after surgery. At 12 h and 24 h after operation, the TNF-a expres sion in the PVA group was significantly higher than that in the control group (P<0.05), but no significant dif fe rence at 7 days after sur gery was observed between the two groups. At 12 h, the HGF expression in the PVA group was similar to that in the con trol group, but significantly higher than in the control group at 24 h (P<0.05). At 24 h, the TGF-b1 expres sion in the PVA group was sig nificantly lower than that in the control group (P <0.05), but no significant difference was found at 48 h after surgery between the two groups. The promo tive effects on the portal vein arterialization at the early stage of liver rege neration were associated with the changes in the expressions of TNF-a, HGF and TGF-b1.

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Section: Real-time Quantitative Rt-pcrmentioning

confidence: 99%

TNF-α, HGF and TGF-β1 are Involved in Liver Regeneration Following Partial Hepatectomy Using Portal Vein Arterializations

Niu¹,

Dong²,

Zhang³

et al. 2012

Journal of Medical Biochemistry

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“…After 48 h, HSC-F cells were harvested. The total viral DNA obtained after the purification procedure (29) was used for SYBR Green real-time PCR assay, as previously described (30), with some modifications. Briefly, primers that recognize specific and highly conserved sequences on the gag region of SIV described by Ui et al (31) were selected.…”

Section: Determination Of Total Number Of Siv Dna Copiesmentioning

confidence: 99%

Targeted Delivery of Immunogen to Primate M Cells with Tetragalloyl Lysine Dendrimer

Misumi

Masuyama

Takamune

et al. 2009

The Journal of Immunology

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Effective uptake of Ags by specialized M cells of gut-associated lymphoid tissues is an important step in inducing efficient immune responses after oral vaccination. Although stable nontoxic small molecule mimetics of lectins, such as synthetic multivalent polygalloyl derivatives, may have potential in murine M cell targeting, it remains unclear whether synthetic multivalent polygalloyl derivatives effectively target nonhuman and human M cells. In this study, we evaluated the ability of a tetragalloyl derivative, the tetragalloyl-d-lysine dendrimer (TGDK), to target M cells in both in vivo nonhuman primate and in vitro human M-like cell culture models. TGDK was efficiently transported from the lumen of the intestinal tract into rhesus Peyer’s patches by M cells and then accumulated in germinal centers. Oral administration of rhesus CCR5-derived cyclopeptide conjugated with TGDK in rhesus macaque resulted in a statistically significant increase in stool IgA response against rhesus CCR5-derived cyclopeptide and induced a neutralizing activity against SIV infection. Furthermore, TGDK was specifically bound to human M-like cells and efficiently transcytosed from the apical side to the basolateral side in the M-like cell model. Thus, the TGDK-mediated vaccine delivery system represents a potential approach for enabling M cell-targeted mucosal vaccines in primates.

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“…A SYBR green-based real-time RT-PCR method using the LightCycler technology was also reported recently [14]. Although each is based on a different scientific principle, they all provide similar results, although it is recommended not to use the results from more than one assay for managing each patient.…”

Section: Monitoring the Efficacy Of Anti-hiv Drug Treatment And Hiv Dmentioning

confidence: 99%

Update on the laboratory diagnosis and monitoring of HIV infection

2005

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In China, the estimated number of HIV infected cases is approaching one million. Although public education has been initiated for awareness and behavioral modification for this devastating infection, better diagnostic methods are needed to identify infected persons and manage infection. Simple and more accurate diagnostic tools have become available, particularly for early detection and to monitor treatment in those who receive anti-retroviral treatment. In this short review, we summarize some of the common and new methodologies that can be used in clinical laboratories, in the field, or in private laboratories. These range from simple antibody tests to more sophistical methods that are used to monitor disease progression and identify drug resistance. These tools can assist physicians, medical practitioners, and laboratory personnel to select suitable diagnostic tools for the diagnosis, blood screening, monitoring of disease progression, and for detection of drug resistance to anti-retroviral therapies.

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Quantitative detection of human immunodeficiency virus type 1 (HIV-1) viral load by SYBR green real-time RT-PCR technique in HIV-1 seropositive patients

Cited by 67 publications

References 27 publications

TNF-α, HGF and TGF-β1 are Involved in Liver Regeneration Following Partial Hepatectomy Using Portal Vein Arterializations

TNF-α, HGF and TGF-β1 are Involved in Liver Regeneration Following Partial Hepatectomy Using Portal Vein Arterializations

Targeted Delivery of Immunogen to Primate M Cells with Tetragalloyl Lysine Dendrimer

Update on the laboratory diagnosis and monitoring of HIV infection

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