Neutrophil-to-lymphocyte ratio, a critical predictor for assessment of disease severity in patients with COVID-19.

INTRODUCTION: Monitoring of laboratory indicators is important for predicting changes in disease severity and clinical outcomes. We aimed to identify the critical predictors that can effectively assess the disease conditions of patients with COVID-19 by analyzing the clinical characteristics and laboratory findings of patients with SARS-CoV-2 infection. METHODS: All consecutive patients (n = 294) with confirmed SARS-CoV-2 infection admitted to the General Hospital of Central Theater Command of the PLA from February 6 to February 21, 2020, were enrolled. These patients were divided into the severe group and the nonsevere group according to disease severity during hospitalization. RESULTS: The median neutrophil-to-lymphocyte ratio (NLR) value of the severe patients was dramatically higher than that of the nonsevere patients (10.4 vs 2.6; P < .001). The NLR value equal to 5 was a boundary value worthy of reference, because more than 80% severe patients had an NLR value greater than 5 and over 80% nonsevere patients had an NLR value less than 5. The NLR value of these COVID-19 patients was positively and respectively correlated with the values of C-reactive protein (R = .5921, P < .001), lactate dehydrogenase (R = .4509, P < .001), procalcitonin (R = .5504, P < .001), fibrinogen (R = .4710, P < .001), and D-dimers (R = .4425, P < .001). However, the NLR value was merely and positively correlated with the interleukin-6 value (R = .3594, P < .05), but had no correlations with the values of interleukin-10, interleukin-4, interleukin-17, interferon-γ, and tumor necrosis factor-α (P > .05). DISCUSSION: Neutrophil-to-lymphocyte ratio is a critical predictor for assessment of disease severity in patients with COVID-19, and it has a close relation with the laboratory indicators related to disease conditions.

[CMV DNA detection in plasma using real-time PCR based on the SYBR-Green I dye method]. / Detección de ADN de CMV en plasma mediante PCR en tiempo real utilizando SYBR-Green I como señal de amplificación.

OBJECTIVE: The aim of this study is to assess a real-time PCR technique on the LightCycler 2.0 with SYBR-Green I detection as compared to another real-time PCR method based on detection with FRET (fluorescence resonance energy transfer) probes for the quantification of CMV DNA. METHODS: The two real-time PCR methods were used to test plasma samples from immunocompromised patients with clinically suspected CMV disease, patients under follow-up without symptoms, and healthy adults. A standard curve for quantitative analysis by the SYBR-Green I method was performed with 10-fold diluted solutions of DNA from the CMV Towne strain (ATCC VR-977) cultured in MRC-5 monolayer. In addition, frozen samples from patients positive for CMV pp65 antigenemia were also analyzed and results compared using the two real time PCR methods. RESULTS: The real-time PCR technique using SYBR-Green I on the LightCycler 2.0 was a highly specific, fast, simple and reliable test to quantify CMV; moreover, it was cost-effective. CONCLUSION: Quantification of CMV DNA in plasma using this sensitive, fast, low-cost method was advantageous for the diagnosis and follow up of patients with opportunistic CMV infection, which are increasingly more frequent in our daily hospital clinical practice.

Human immunodeficiency virus load. Quantitative assessment in semen from seropositive individuals and in spiked seminal plasma.

OBJECTIVE: To establish criteria for the quantitation of the human immunodeficiency virus (HIV) in seminal plasma, seminal cells and the whole semen of HIV-infected individuals. The reverse transcription polymerase chain reaction (RT-PCR), DNA-PCR and semen HIV culture assays were standardized by testing seminal plasma spiked separately with serial dilutions of cell-free and cell-associated HIV stocks of known titers. The standardized assays were then used to assess the quantity of virus in the freshly collected seminal cells and seminal plasma. RESULTS: Analysis of freshly collected peripheral blood mononuclear cells (PBMCs) and paired semen from HIV-seropositive men who had received antiviral drugs and/or immunemodulators indicated that HIV could be isolated from 42 of 55 (76%) samples of peripheral blood mononuclear cells (PBMCs) and 13 of 55 (24%) samples of ejaculates. Since no semen sample was culture positive in the absence of culturable HIV in PBMCs of the same individual, RT-PCR was 5-125 times more sensitive than cell cultures for the quantitation of HIV spiked in seminal plasma, freshly collected seminal fluid and whole semen. Further, HIV-RNA was detected in samples containing higher dilutions of virus from which HIV was not isolated by culture. CONCLUSION: We conclude that cell-free HIV is present in excess of the culturable virus in all specimens tested and that the high sensitivity of HIV-RNA detection is useful for quantitation of the virus directly in seminal fluid, seminal cells and whole semen.