A new generic real-time PCR kit for hepatitis delta RNA viral load quantification

The aim of this study was to investigate the persistence of SARS-CoV-2 in post-mortem swabs of subjects who died from SARS-CoV-2 infection. The presence of the virus was evaluated post-mortem from airways of 27 SARS-CoV-2 positive patients at three different time points (T1 2 h; T2 12 h; T3 24 h) by real-time PCR. Detection of antibodies to SARS-CoV-2 was performed by Maglumi 2019-nCoV IgM/IgG chemiluminescence assay. SARS-CoV-2 viral RNA was still detectable in 70.3% of cases within 2 h after death and in 66,6% of cases up to 24 h after death. Our data showed an increase of the viral load in 78,6% of positive individuals 24 h post-mortem (T3) in comparison to that evaluated 2 h after death (T1). Noteworthy, we detected a positive T3 post-mortem swab (24 h after death) from 4 subjects who were negative at T1 (2 h after death). The results of our study may have an important value in the management of deceased subjects not only with a suspected or confirmed diagnosis of SARS-CoV-2, but also for unspecified causes and in the absence of clinical documentation or medical assistance.

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Development of a standardized real time PCR for Torque teno viruses (TTV) viral load detection and quantification: A new tool for immune monitoring

Journal of Clinical Virology ◽

10.1016/j.jcv.2018.06.010 ◽

2018 ◽

Vol 105 ◽

pp. 118-127 ◽

Cited By ~ 15

Author(s):

Dorian Kulifaj ◽

Bénédicte Durgueil-Lariviere ◽

Faustine Meynier ◽

Eliza Munteanu ◽

Nicolas Pichon ◽

...

Keyword(s):

Viral Load ◽

Real Time ◽

Real Time Pcr ◽

Immune Monitoring ◽

Load Detection ◽

Detection And Quantification

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REAL-TIME PCR PER IL VIRAL LOAD DI CITOMEGALOVIRUS NEL TRAPIANTO D’ORGANO: CONFRONTO CON ANTIGENEMIA E PCR END-POINT.

Microbiologia Medica ◽

10.4081/mm.2004.3765 ◽

2004 ◽

Vol 19 (2) ◽

Author(s):

T. Zaccaria ◽

M. Enrietto ◽

F. Pittaluga ◽

V. Ghisetti ◽

G. Marchiaro

Keyword(s):

Viral Load ◽

Real Time ◽

Real Time Pcr ◽

End Point

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Salivary and Nasal Detection of the SARS-CoV-2 Virus After Antiviral Mouthrinses (BBCovid): A structured summary of a study protocol for a randomised controlled trial

Trials ◽

10.1186/s13063-020-04846-6 ◽

2020 ◽

Vol 21 (1) ◽

Author(s):

Florence Carrouel ◽

Stéphane Viennot ◽

Martine Valette ◽

Jean-Marie Cohen ◽

Claude Dussart ◽

...

Keyword(s):

Viral Load ◽

Randomised Controlled Trial ◽

Real Time ◽

Outcome Measures ◽

Real Time Pcr ◽

Controlled Trial ◽

Full Protocol ◽

Pcr Assays ◽

Randomised Controlled ◽

Hospital Center

Abstract Objectives - To describe the evolution of the SARS-CoV-2 salivary viral load of patients infected with Covid-19, performing 7 days of tri-daily mouthwashes with and without antivirals. - To compare the evolution of the SARS-CoV-2 nasal and salivary viral load according to the presence or absence of antivirals in the mouthwash. Trial design This is a multi-center, randomised controlled trial (RCT) with two parallel arms (1:1 ratio). Participants Inclusion criteria - Age: 18-85 years old - Clinical diagnosis of Covid-19 infection - Clinical signs have been present for less than 8 days - Virological confirmation - Understanding and acceptance of the trial - Written agreement to participate in the trial Exclusion criteria - Pregnancy, breastfeeding, inability to comply with protocol, lack of written agreement - Patients using mouthwash on a regular basis (more than once a week) - Patient at risk of infectious endocarditis - Patients unable to answer questions - Uncooperative patient The clinical trial is being conducted with the collaboration of three French hospital centers: Hospital Center Emile Roux (Le Puy en Velay, France), Clinic of the Protestant Infirmary (Lyon, France) and Intercommunal Hospital Center (Mont de Marsan, France). Intervention and comparator Eligible participants will be allocated to one of the two study groups. Intervention group: patients perform a tri-daily mouthwash with mouthwash containing antivirals (β-cyclodextrin and Citrox®) for a period of 7 days. Control group: patients perform a tri-daily mouthwash with a placebo mouthwash for a period of 7 days. Main outcomes Primary Outcome Measures: Change from Baseline amount of SARS-CoV-2 in salivary samples at 4 and 9 hours, 1, 2, 3, 4, 5 and 6 days. Real-time PCR assays are performed to assess salivary SARS-CoV 2 viral load. Secondary Outcome Measures: Change from Baseline amount of SARS-CoV-2 virus in nasal samples at 6 days. Real-time PCR assays are performed to assess nasal SARS-CoV-2 viral load. Randomisation Participants meeting all eligibility requirements are allocated to one of the two study arms (mouthwash with β-cyclodextrin and Citrox® or mouthwash without β-cyclodextrin and Citrox®) in a 1:1 ratio using simple randomisation with computer generated random numbers. Blinding (masking) Participants, doctors and nurses caring for participants, laboratory technicians and investigators assessing the outcomes will be blinded to group assignment. Numbers to be randomised (sample size) Both the intervention and control groups will be composed of 103 participants, so the study will include a total of 206 participants. Trial Status The current protocol version is 6, August 4th, 2020. Recruitment began on April 6, 2020 and is anticipated to be complete by April 5, 2021. As of October 2, 2020, forty-two participants have been included. Trial registration This trial was registered on 20 April 2020 at www.clinicaltrials.gov with the number NCT04352959. Full protocol The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.” The study protocol has been reported in accordance with the Standard Protocol Items: Recommendations for Clinical Interventional Trials (SPIRIT) guidelines (Additional file 2).”

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Evaluation of the HBV Genotype, Viral Load and Antivirus Drug Resistance Mutation in Tay Ninh Hospital, Vietnam by Real-Time PCR

IFMBE Proceedings - 7th International Conference on the Development of Biomedical Engineering in Vietnam (BME7) ◽

10.1007/978-981-13-5859-3_103 ◽

2019 ◽

pp. 605-609

Author(s):

Thuan Duc Lao ◽

Hung Chi Lieu ◽

Thuy Thanh Thi Ho ◽

Thuy Ai Huyen Le

Keyword(s):

Drug Resistance ◽

Viral Load ◽

Real Time ◽

Real Time Pcr ◽

Resistance Mutation ◽

Drug Resistance Mutation ◽

Hbv Genotype

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Weekly High-Dose Cidofovir Is an Effective Treatment for Symptomatic BK Virus (BKV)-Associated Hemorrhagic Cystitis (HC) after Allogeneic Stem Cell Transplantation (Allo-SCT).

Blood ◽

10.1182/blood.v108.11.5306.5306 ◽

2006 ◽

Vol 108 (11) ◽

pp. 5306-5306

Author(s):

Ioannis Baltathakis ◽

Michael Michael ◽

Eirini Grispou ◽

Georgia Kourti ◽

Maria Bouzani ◽

...

Keyword(s):

Viral Load ◽

Real Time ◽

Median Time ◽

Real Time Pcr ◽

Dose Schedule ◽

Bk Virus ◽

High Dose ◽

Complete Suppression ◽

Cmv Infection ◽

Papillary Necrosis

Abstract A proportion of cases of HC after allo-SCT are associated with infection from BKV. Supportive treatment is not sufficient for the relief of symptoms in many patients with BKV-associated HC. Cidofovir is an antiviral agent active against BKV. Its use is limited, however, due to the reported risk of renal failure in patients with cytomegalovirus (CMV) infection. The experience with cidofovir in BKV-associated HC is anecdotal, and the optimal dose schedule of the drug has not been determined. We administered weekly high-dose cidofovir in 5 patients for the treatment of 6 episodes of symptomatic BKV-associated HC, which were refractory to first-line therapy with hydration and continuous bladder irrigation. One patient was treated twice with cidofovir for 2 temporally separated episodes of HC, combined with papillary necrosis and ureteritis respectively. All 5 patients were males, aged 21–44 years, and had received grafts from matched related (n=2), matched unrelated (n=1) or haploidentical (n=1) donors. The median time from transplantation to diagnosis of HC was 61.5 (range, 26–303) days. In all cases, the development of HC was associated with severe immunosuppression due to profound lymphopenia or treatment with steroids or antithymocyte globulin for GVHD. BKV-associated HC was confirmed by detection of BKV in the urine by real-time PCR (1.5 × 107–5.5 × 109 copies/ml). In 2 of the 6 episodes, the patients also developed BKV viremia (5.0–9.9 × 103 copies/ml). The median time from the manifestation of symptoms to initiation of cidofovir was 12 (range, 4–53) days. Cidofovir was started at a dose of 2.5 mg/kg/week intravenously over 1 hour. If renal function remained stable, subsequent doses of 5 mg/kg were given at weekly intervals for a total of 4–6 doses. Oral probenecid and intravenous hydration were administered in combination with cidofovir. No patient developed renal toxicity (a rise in serum creatinine of at least 1.5 × baseline or proteinuria). In 5 of the 6 cases, clinical improvement of patients was observed, with a median time to response of 5 (range, 3–8) days after the first dose of cidofovir. Complete resolution of symptoms of HC was achieved in these 5 cases after 4 to 6 doses of cidofovir. BKV viral load was monitored weekly in urine (and blood) during therapy. Clinical responses did not correlate with a reduction in viral load in urine. However, in the 2 cases with BKV viremia, real-time PCR became negative after treatment with cidofovir. Concomitant CMV infection was detected in 5 of the 6 episodes of BKV-associated HC, and in all but one, complete suppression of CMV reactivation was achieved by cidofovir. Four of the 5 patients are currently alive and free of symptoms of HC. In conclusion, treatment of BKV-associated HC with cidofovir at the dose of 5 mg/kg/week seems to be safe and may result in prompt and sustained relief of the debilitating symptoms of this condition. Additional manifestations of BKV-associated disease (papillary necrosis and ureteritis) and concurrent CMV infection can be managed successfully with this dose schedule. To our knowledge, these 6 cases comprise the largest reported experience of treatment of BKV-associated HC with cidofovir, albeit the encouraging results need to be substantiated by further trials.

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