scholarly journals Appraisal of a Simple and Effective RT-qPCR Assay for Evaluating the Reverse Transcriptase Activity in Blood Samples from HIV-1 Patients

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1047
Author(s):  
Beatrice Macchi ◽  
Caterina Frezza ◽  
Francesca Marino-Merlo ◽  
Antonella Minutolo ◽  
Valeria Stefanizzi ◽  
...  

Testing HIV-1 RNA in plasma by PCR is universally accepted as the ultimate standard to confirm diagnosis of HIV-1 infection and to monitor viral load in patients under treatment. However, in some cases, this assay could either underestimate or overestimate the replication capacity of a circulating or latent virus. In the present study, we performed the assessment of evaluating the HIV-1 reverse transcriptase (RT) activity by means of a new assay for the functional screening of the status of HIV-1 patients. To this purpose, we utilized, for the first time on blood samples, an adapted version of a real-time RT quantitative PCR assay, utilized to evaluate the HIV-1-RT inhibitory activity of compounds. The study analyzed blood samples from 28 HIV-1-infected patients, exhibiting a wide range of viremia and immunological values. Results demonstrated that plasma HIV-1 RT levels, expressed as cycle threshold values obtained with the assay under appraisal, were inversely and highly significantly correlated with the plasma HIV-1-RNA levels of the patients. Thus, an HIV-1 RT quantitative PCR assay was created which we describe in this study, and it may be considered as a promising basis for an additional tool capable of furnishing information on the functional virological status of HIV-1-infected patients.

Author(s):  
Alexandre Alanio ◽  
Maud Gits-Muselli ◽  
Fanny Lanternier ◽  
Aude Sturny-Leclère ◽  
Marion Benazra ◽  
...  

2006 ◽  
Vol 133 (2) ◽  
pp. 185-194 ◽  
Author(s):  
N.M. van Maarseveen ◽  
M.C.D.G. Huigen ◽  
D. de Jong ◽  
A.M. Smits ◽  
C.A.B. Boucher ◽  
...  

2003 ◽  
Vol 47 (5) ◽  
pp. 1665-1671 ◽  
Author(s):  
Florence Fenollar ◽  
Max Maurin ◽  
Didier Raoult

ABSTRACT Wolbachia spp. are strict intracellular bacteria that infect a wide range of arthropods and filarial nematodes. Filarial nematodes are important causes of human diseases. There is increasing evidence that Wolbachia spp. influence important functions in the biology of the hosts, specifically, infertility. Preliminary experiments with humans and animals have suggested that antibiotics with activity against Wolbachia may help to treat filariasis. In this study, we determined using a real-time quantitative PCR assay the growth kinetics of a strain of Wolbachia pipientis from a mosquito grown in Aa23 cells. The doubling time was estimated to be 14 h. We then determined the susceptibilities of this strain to 13 antibiotics by two methods: an immunofluorescent-antibody test and a real-time quantitative PCR assay. Both techniques gave similar results. Doxycycline and rifampin were the most effective compounds, with MICs of 0.125 and 0.06 to 0.125 μg/ml, respectively. Fluoroquinolones were less effective, with MICs of 2 to 4 μg/ml for ciprofloxacin, 2 μg/ml for ofloxacin, and 1 μg/ml for levofloxacin. β-Lactams (penicillin G, amoxicillin, ceftriaxone) were not effective at concentrations up to 128 μg/ml. The MIC of erythromycin was >32 μg/ml, whereas that of telithromycin was 8 μg/ml. Other antibiotic compounds were bacteriostatic only at high concentrations, including gentamicin, co-trimoxazole, and thiamphenicol. The real-time PCR assay was a convenient and reliable technique for determination of the antibiotic susceptibilities of Wolbachia. It may help in the future to simplify antibiotic susceptibility testing of strict intracellular pathogens.


Sign in / Sign up

Export Citation Format

Share Document