Sensibilidad y especificidad de la técnica de detección de DNA de citomegalovirus en la sangre seca de la prueba de detección precoz neonatal (prueba del talón) mediante PCR en los pacientes afectos de citomegalovirus congénito

La infección congénita por citomegalovirus (CMVc) está considerada la infección congénita más frecuente de los países desarrollados, estimándose su prevalencia en un 0,7% (entre 0,3 y 1,3%). Actualmente, ningún país del mundo realiza cribado universal de CMVc. Para su diagnóstico se debe confirmar la presencia del virus en cualquier fluido corporal (el más frecuentemente usado es la orina) en las dos o tres primeras semanas de vida, pues pasado ese periodo la adquisición puede ser postnatal. Para diferenciar retrospectivamente entre la adquisición pre y postnatal se determina la presencia de DNA del virus mediante PCR en la sangre seca de la prueba de cribado metabólico neonatal. Existen múltiples estudios publicados sobre la sensibilidad de estas técnicas, sólo uno en nuestro país, con sensibilidades entre el 28 y el 100%. El estudio piloto realizado por nuestro grupo en 2014 sobre 14 pacientes mostró una sensibilidad menor (50%) a la descrita en la mayoría de estudios. En este trabajo multicéntrico se ha estudiado la sensibilidad y especificidad de una técnica comercial de rt-PCR (rt-PCR (RealStar CMV®, Altona, Hamburg, Germany)) entre los pacientes del Registro Estatal De Infección Congénita por CMV (REDICCMV), así como los factores relacionados con el resultado. En este trabajo se incluyeron 184 individuos (103 pacientes y 81 controles). Cincuenta y ocho pacientes y dos controles presentaron resultado positivo en la PCR a CMV en sangre seca, lo que equivale a una sensibilidad de la prueba del 56% (95%CI 47-65%), especificidad del 98% (91-99%), razón de probabilidad positiva del 22,81 (5,74-90,58) y razón de probabilidad negativa del 0,45 (0,36-0,60). Al analizar las variables clínicas en relación al resultado de la PCR se observa que únicamente la carga viral al nacimiento tiene relación con la misma: cargas virales más bajas al nacimiento tienen con más frecuencia rt-PCRs negativas en la sangre seca. Sin embargo, al estudiar la correlación entre la carga viral en sangre al nacimiento y en sangre seca (técnica semicuantitativa), no se ha observado ninguna correlación (se ha podido estudiar en 51 pacientes). Como conclusión podemos decir que, con la técnica diagnóstica y metodología utilizadas, ante un resultado negativo de la rt-PCR en la muestra de sangre seca de la prueba de cribado metabólico no podemos descartar infección congénita por CMV, sobre todo en los niños con cargas virales más bajas al nacimiento. Por el contrario, un resultado positivo tiene una elevada especificidad para el diagnóstico de confirmación de CMVc. Es necesario disponer de una técnica definida en cuanto a muestra, extracción, procesamiento e interpretación que permita el diagnóstico retrospectivo de la infección por CMVc cuando ello sea necesario.Congenital cytomegalovirus (cCMV) is known to be the most common cause of congenital viral infection of the developed countries, its prevalence among 0,7% (between 0,3 and 1,3%). Currently, there is no country worldwide with a universal screening program. The diagnosis of a congenital infection requires the confirmation of the virus in any body fluid (urine is the most commonly used) within the first two or three weeks of life. Afterwards, the detection of CMV can be due to postnatal infection. If we need to make a retrospective diagnosis of congenital or postnatal infection when we suspect the infection after the first two or three weeks of life, the DNA presence of the virus is checked by a PCR technique in dried blood spots (DBS) from the metabolic screening program. There have been many published studies about this topic, only one in Spain, with sensitivities ranging from 28% until 100%. The pilot study made by our group with 14 patients showed a lower sensitivity (50%), compared with those reported in most of the previously published studies. In this multicentric study the sensitivity and specificitiy of a commercial rt-PCR (rt-PCR (RealStar CMV®, Altona, Hamburg, Germany)) has been assessed in the Spanish Registry of Congenital CMV Infection (Registro Estatal De Infección Congénita por CMV (REDICCMV)), as well as the relation between the result and clinical variables. REDICCMV is an online registry including all those cCMV patients from hospitals around the country included in the project. One hundred and eighty-four subjects were included (103 patients and 81 controls). Fifty-eight samples from the patient group in contrast to two among the control samples tested positive by CMV DNA rt-PCR, leading to a sensitivity of 56% (95%CI 47-65%), specificity 98% (91-99%), positive likelihood ratio 22.81 (5.74-90.58) and negative likelihood ratio 0.45 (0.36-0.60). The only variable significantly associated with negative CMV DNA results on DBS testing was the plasma viral load at birth (bPVL): those patients with lower bPVL tested negative more frequently than those with higher bPVL. However, no correlation between bPVL and viral load in DBS (semi quantitative technique) was found in those patients that could be checked for both viral loads (51). In conclusion, with our technique and used methodology, a negative rt-PCR DBS result does not rule out congenital CMV infection, especially in patients with low viremia at birth. In contrast, a positive result has a high specificity for the retrospective diagnosis of cCMV infection. There is a need of an available technique regarding sampling, extraction, processing and result interpretation for the retrospective diagnosis of cCMV when necessary

Sensibilidad y especificidad de la técnica de detección de DNA de citomegalovirus en la sangre seca de la prueba de detección precoz neonatal (prueba del talón) mediante PCR en los pacientes afectos de citomegalovirus congénito /

La infección congénita por citomegalovirus (CMVc) está considerada la infección congénita más frecuente de los países desarrollados, estimándose su prevalencia en un 0,7% (entre 0,3 y 1,3%). Actualmente, ningún país del mundo realiza cribado universal de CMVc. Para su diagnóstico se debe confirmar la presencia del virus en cualquier fluido corporal (el más frecuentemente usado es la orina) en las dos o tres primeras semanas de vida, pues pasado ese periodo la adquisición puede ser postnatal. Para diferenciar retrospectivamente entre la adquisición pre y postnatal se determina la presencia de DNA del virus mediante PCR en la sangre seca de la prueba de cribado metabólico neonatal. Existen múltiples estudios publicados sobre la sensibilidad de estas técnicas, sólo uno en nuestro país, con sensibilidades entre el 28 y el 100%. El estudio piloto realizado por nuestro grupo en 2014 sobre 14 pacientes mostró una sensibilidad menor (50%) a la descrita en la mayoría de estudios. En este trabajo multicéntrico se ha estudiado la sensibilidad y especificidad de una técnica comercial de rt-PCR (rt-PCR (RealStar CMV®, Altona, Hamburg, Germany)) entre los pacientes del Registro Estatal De Infección Congénita por CMV (REDICCMV), así como los factores relacionados con el resultado. En este trabajo se incluyeron 184 individuos (103 pacientes y 81 controles). Cincuenta y ocho pacientes y dos controles presentaron resultado positivo en la PCR a CMV en sangre seca, lo que equivale a una sensibilidad de la prueba del 56% (95%CI 47-65%), especificidad del 98% (91-99%), razón de probabilidad positiva del 22,81 (5,74-90,58) y razón de probabilidad negativa del 0,45 (0,36-0,60). Al analizar las variables clínicas en relación al resultado de la PCR se observa que únicamente la carga viral al nacimiento tiene relación con la misma: cargas virales más bajas al nacimiento tienen con más frecuencia rt-PCRs negativas en la sangre seca. Sin embargo, al estudiar la correlación entre la carga viral en sangre al nacimiento y en sangre seca (técnica semicuantitativa), no se ha observado ninguna correlación (se ha podido estudiar en 51 pacientes). Como conclusión podemos decir que, con la técnica diagnóstica y metodología utilizadas, ante un resultado negativo de la rt-PCR en la muestra de sangre seca de la prueba de cribado metabólico no podemos descartar infección congénita por CMV, sobre todo en los niños con cargas virales más bajas al nacimiento. Por el contrario, un resultado positivo tiene una elevada especificidad para el diagnóstico de confirmación de CMVc. Es necesario disponer de una técnica definida en cuanto a muestra, extracción, procesamiento e interpretación que permita el diagnóstico retrospectivo de la infección por CMVc cuando ello sea necesario.Congenital cytomegalovirus (cCMV) is known to be the most common cause of congenital viral infection of the developed countries, its prevalence among 0,7% (between 0,3 and 1,3%). Currently, there is no country worldwide with a universal screening program. The diagnosis of a congenital infection requires the confirmation of the virus in any body fluid (urine is the most commonly used) within the first two or three weeks of life. Afterwards, the detection of CMV can be due to postnatal infection. If we need to make a retrospective diagnosis of congenital or postnatal infection when we suspect the infection after the first two or three weeks of life, the DNA presence of the virus is checked by a PCR technique in dried blood spots (DBS) from the metabolic screening program. There have been many published studies about this topic, only one in Spain, with sensitivities ranging from 28% until 100%. The pilot study made by our group with 14 patients showed a lower sensitivity (50%), compared with those reported in most of the previously published studies. In this multicentric study the sensitivity and specificitiy of a commercial rt-PCR (rt-PCR (RealStar CMV®, Altona, Hamburg, Germany)) has been assessed in the Spanish Registry of Congenital CMV Infection (Registro Estatal De Infección Congénita por CMV (REDICCMV)), as well as the relation between the result and clinical variables. REDICCMV is an online registry including all those cCMV patients from hospitals around the country included in the project. One hundred and eighty-four subjects were included (103 patients and 81 controls). Fifty-eight samples from the patient group in contrast to two among the control samples tested positive by CMV DNA rt-PCR, leading to a sensitivity of 56% (95%CI 47-65%), specificity 98% (91-99%), positive likelihood ratio 22.81 (5.74-90.58) and negative likelihood ratio 0.45 (0.36-0.60). The only variable significantly associated with negative CMV DNA results on DBS testing was the plasma viral load at birth (bPVL): those patients with lower bPVL tested negative more frequently than those with higher bPVL. However, no correlation between bPVL and viral load in DBS (semi quantitative technique) was found in those patients that could be checked for both viral loads (51). In conclusion, with our technique and used methodology, a negative rt-PCR DBS result does not rule out congenital CMV infection, especially in patients with low viremia at birth. In contrast, a positive result has a high specificity for the retrospective diagnosis of cCMV infection. There is a need of an available technique regarding sampling, extraction, processing and result interpretation for the retrospective diagnosis of cCMV when necessary

Growth Patterns in Children with Congenital Cytomegalovirus Infection

Background: Congenital cytomegalovirus infection (CMVc) affects 0.7%-6% of recent births. Among its clinical manifestations are low weight and length at birth. Objective: Describe the growth patterns of children with CMVc in their early years. Methods: Observational, multicenter study of patients with CMVc. Anthropometric data were collected during the first 2 years of life and compared with World Health Organization standards. Results: Anthropometric characteristics of 383 children with CMVc were studied, of which 198 (51%) were symptomatic at birth. At birth, 9% were small for gestational age (SGA) in terms of their weight and length and 17% had microcephaly. At 24 ± 3 months, 10% had a weight and length ≤2 SD, and 13% a head circumference ≤2 SD. Of those who were SGA at birth, at 24 ± 3 months >20% remained at ≤2 SD of their weight and length. Conversely, 75% of children with low weight or length at 24 ± 3 had not been SGA at birth. 20% of infants with microcephaly at birth remained with microcephaly, and 10% of those without microcephaly developed it at 24 ± 3 months. The average growth rate in length and weight was normal. Patients who were symptomatic at birth, premature and with motor and neurocognitive impairment had a significantly higher risk of low weight and length at 24 ± 3 months. Conclusion: Around 10% of children with CMVc are at ≤2 SD in weight, length and head circumference at 24 ± 3 months. The lack of adequate growth is associated with symptoms at birth, prematurity and motor and neurocognitive impairment. Growth impairment could be incorporated into the symptomatic spectrum of CMVc.Sin financiación3.164 JCR (2020) Q3, 56/92 Infectious Diseases1.269 SJR (2020) Q1, 69/292 Infectious DiseasesNo data IDR 2019UE

Cytomegalovirus DNA Detection by Polymerase Chain Reaction in Cerebrospinal Fluid of Infants With Congenital Infection: Associations With Clinical Evaluation at Birth and Implications for Follow-up

BACKGROUND: DNA detection of human cytomegalovirus (hCMV) in cerebrospinal fluid (CSF) by polymerase chain reaction (PCR) is a marker of central nervous system (CNS) involvement in congenital hCMV infection (cCMV), but its prognostic value is unknown. METHODS: A multicenter, retrospective study was performed using the Spanish Congenital Cytomegalovirus Infection Database (REDICCMV; http://www.cmvcongenito.es). Newborns with cCMV and a lumbar puncture performed were included and classified according to their hCMV-PCR in CSF result (positive/negative). Clinical characteristics, neuroimaging abnormalities, plasma viral load, and audiological and neurological outcomes of both groups were compared. RESULTS: A total of 136 neonates were included in the study: 21 (15.4%) with positive CSF hCMV-PCR and 115 (84.6%) with negative results. Seventeen patients (81%) in the positive group were symptomatic at birth compared with 52.2% of infants in the negative group (odds ratio [OR], 3.86; 95% confidence interval [CI], 1.28-14.1; P = .01). Only 4 asymptomatic newborns (6.8%) had a positive CSF hCMV-PCR. There were no differences between groups regarding the rate of microcephaly, neuroimaging abnormalities, neurological sequelae at 6 months of age, or plasma viral load. Sensorineural hearing loss (SNHL) at birth was associated with a positive CSF hCMV-PCR result (OR, 3.49; 95% CI, 1.08-11.27; P = .04), although no association was found at 6 months of age. CONCLUSIONS: A positive hCMV-PCR result in CSF is associated with symptomatic cCMV and SNHL at birth. However, no differences in neuroimaging studies, plasma viral load, or outcomes at 6 months were found. These results suggest that hCMV-PCR in CSF may not be a useful prognostic marker in cCMV.Sin financiación9.117 JCR (2017) Q1, 11/155 Immunology, 3/88 Infectious Diseases, 11/125 MicrobiologyUE