NASBATM isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection

Kievits, Tim; Gemen, Bob van; Strijp, Dianne van; Schukkink, R; Dircks, Mariët; Adriaanse, Henriëtte; Malek, L T; Sooknanan, Roy; Lens, P.

doi:10.1016/0166-0934(91)90069-c

Cited by 405 publications

(209 citation statements)

References 17 publications

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“…Real-time PCR assay has been useful to study the transmission and development of this viral infection in juvenile [56]. Another useful method is the nucleic acid sequence based amplification (NASBA) [27] which is an isothermal method for nucleic acid amplification that is particularly suited to RNA targets [68]. The method amplifies a target-specific product through oligonucleotide primers and the co-ordinated activity of 3 enzymes: reverse transcriptase, RNase H, and T7 RNA polymerase.…”

Section: Microscopymentioning

confidence: 99%

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

et al. 2012

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Section: Microscopymentioning

confidence: 99%

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

et al. 2012

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“…18,19 In this study, we monitored and analyzed CMV activities using an NASBA technique, which is highly suited for the amplification of RNA sequences. [7][8][9] We amplified the ␤2.7 transcripts of the CMV, which is detected concomitantly with active viral replication. 10 Thus, viral activation is indicated by detecting mRNA not viral DNA.…”

Section: Discussionmentioning

confidence: 99%

Foscarnet therapy for ganciclovir-resistant cytomegalovirus retinitis after stem cell transplantation: effective monitoring of CMV infection by quantitative analysis of CMV mRNA

Ohta

Matsuda

Tokimasa

et al. 2001

Bone Marrow Transplant

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Summary:We report three pediatric patients with ganciclovirresistant cytomegalovirus (CMV) retinitis who were successfully treated with foscarnet. The patients were recipients of hematopoietic stem cell transplantation (SCT) from HLA-mismatched donors. Because these patients had developed or experienced progressive CMV retinitis during ganciclovir therapy, they received foscarnet therapy at 60 mg/kg every 8 h. Their retinitis resolved promptly after initiating foscarnet therapy, suggesting foscarnet's effectiveness in treating ganciclovirresistant CMV infection. The amount of CMV mRNA was quantitatively measured using an NASBA technique, which amplified the ␤2.7 transcripts specific for CMV replication. This technique was useful for monitoring disease activity in a more rapid and sensitive manner than the PCR assay for CMV DNA. Bone Marrow Transplantation (2001) 27, 1141-1145. Keywords: cytomegalovirus; ganciclovir; foscarnet; resistance; hematopoietic stem cell transplantation; NASBA Cytomegalovirus (CMV) infections are a major cause of morbidity and mortality among immunocompromised patients, especially recipients of allogeneic hematopoietic stem cell transplantation (SCT) and patients with AIDS. The measurement of CD4 + T cell count is crucial in patients at high risk of CMV retinitis; CMV retinitis develops when CD4 + T cell counts have decreased to less than 100/ l. A combination of ganciclovir and intravenous immunoglobulin is effective in treating CMV diseases. However, the development of resistance to ganciclovir has been reported. [1][2][3][4] Foscarnet (trisodium phosphonoformate, PFA), a pyrophosphate analog, 5 has been used in patients with AIDS, and allogeneic SCT recipients. Because the viral inhibitory mechanism of foscarnet is distinct from that of ganciclovir, foscarnet might be useful in patients with ganciclovir-resistant CMV infection. 2,4,6 It usually takes 2 weeks for retinitis to heal on intravenous ganciclovir or foscarnet.We report the clinical and virologic results of foscarnet therapy in three recipients who underwent SCT and developed ganciclovir-resistant CMV retinitis. CMV activities were monitored with a nucleic acid sequence-based amplification (NASBA) technique, 7-9 which amplified the ␤2.7 transcripts specific for CMV replication. 10 Thus, detecting CMV mRNA indicates active viral replication, which is not proven when only viral DNA is detected. Patients and methods PatientsThis study included three patients with acute leukemia who received an allogeneic SCT between 1997 and 1999. The ages of patients 1, 2 and 3 were 10 years, 16 years and 10 months, respectively, and CMV retinitis developed 104, 103 and 42 days after SCT, respectively. The pre-transplant CMV serological status of donors and recipients was positive. Diagnosis of CMV retinitisOphthalmologic evaluation: Patients were evaluated by experienced ophthalmologists using indirect ophthalmoscopy at least every 4 weeks after SCT and whenever virologic examination proved positive. Criteria for the diagnosis of CMV retinitis...

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“…HCV RNA is usually detected in serum by sensitive polymerase chain reaction (PCR)-based techniques and has become a useful tool for diagnosis and monitoring. Besides methods for qualitative detection of viremia, a number of procedures to quantify serum HCV RNA have been developed, including end-point dilution PCR, 1 competitive PCR, 2,3 isothermal nucleic acid amplification, 4 and signal-amplification branched DNA. 5 Routine use of these techniques in a wide clinical setting is hampered by problems of specificity and sensitivity, lack of reproducibility, poor standardization, and high cost.…”

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confidence: 99%

Hepatitis C virus RNA profiles in chronically infected individuals: Do they relate to disease activity?

et al. 1999

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Fluctuations of hepatitis C virus (HCV)-RNA serum levels were monitored in a multicenter study in 76 chronic HCV carriers who had been followed longitudinally without receiving antiviral therapy to assess their relation with the course of liver disease activity. Forty-four patients had normal transaminases over more than 2 years, while 32 additional patients had fluctuating levels. Viral load was measured in serial serum samples prospectively collected for 10 to 12 months in 54 patients and in sera stored yearly up to 8 years in an additional 22 patients. In patients tested monthly, a lesser extent of fluctuations was detected in cases with constantly normal transaminases as compared with those with fluctuating transaminases. In the former group, the mean difference between maximum and minimum values observed in each individual patient was 0.7 Log, while in the latter group, it was 1.3 Log (P ‫؍‬ .0004). Most of these patients experienced, on average, three peaks of viremia over 1 year. The range of variation observed upon yearly testing was between 0.2 and 2.2 Log and did not reach statistical significance between the two groups. In conclusion, a careful viral replication profile can be achieved only by monthly testing, because longer time intervals could miss viremia fluctuations. HCV-RNA levels are more stable in asymptomatic HCV carriers than in patients with biochemical activity of liver disease. (HEPATOLOGY 1999;29: 585-589.)Chronic infection with hepatitis C virus (HCV) is characterized by persistent viremia. HCV RNA is usually detected in serum by sensitive polymerase chain reaction (PCR)-based techniques and has become a useful tool for diagnosis and monitoring. Besides methods for qualitative detection of viremia, a number of procedures to quantify serum HCV RNA have been developed, including end-point dilution PCR, 1 competitive PCR, 2,3 isothermal nucleic acid amplification, 4 and signal-amplification branched DNA. 5 Routine use of these techniques in a wide clinical setting is hampered by problems of specificity and sensitivity, lack of reproducibility, poor standardization, and high cost. 6 Recently, methods for quantitative assessment of HCV-RNA levels have become commercially available and are being extensively evaluated in clinical studies, particularly in patients treated with interferons and antivirals. 7 Little is known about viral kinetics in untreated patients, because most evaluations of the level of viremia are based on single determinations. If wide spontaneous fluctuations occur, they could mimic treatment-induced effects or lead to under-or overestimate baseline values. In a few published studies, different time intervals have been evaluated and discordant conclusions have been reported. Either trivial or consistent differences of viral load over time have been described after observation periods ranging from days to months. [8][9][10][11][12] To assess the spontaneous behavior of serum HCV-RNA levels over a reasonable length of time upon close observation, we monitored chronic HCV car...

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NASBATM isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection

Cited by 405 publications

References 17 publications

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

Foscarnet therapy for ganciclovir-resistant cytomegalovirus retinitis after stem cell transplantation: effective monitoring of CMV infection by quantitative analysis of CMV mRNA

Hepatitis C virus RNA profiles in chronically infected individuals: Do they relate to disease activity?

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