Determination of HPV type 16 and 18 viral load in cervical smears of women referred to colposcopy

Carcopino, Xavier; Henry, Mireille; Benmoura, D.; Fallabregues, A.S.; Richet, Hervé; Boubli, L.; Tamalet, Catherine

doi:10.1002/jmv.20673

Cited by 62 publications

(66 citation statements)

References 34 publications

(26 reference statements)

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“…Many previous studies have found a positive association between viral load and disease severity, which has led to the idea of using viral load as a biomarker to determine the status of cervical disease or to predict its progression (4,11,13,19,23,24,28,30,31). However, at the same time, contradictory data have also been generated (1,15,26,34).…”

Section: Discussionmentioning

confidence: 99%

“…As the disease worsens, the virus becomes integrated, which is a genome form not capable of self-replicating, and therefore the mechanism for maintaining a high viral load is uncertain. Many studies that attempted to examine viral load associations did not analyze the physical status of the viral genome (4,12,13,18,28,29). In addition, some studies have used a method that measured the total viral load of a mixture of HPV types (13,18,28,29).…”

Section: Discussionmentioning

confidence: 99%

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Analysis of Human Papillomavirus Type 18 Load and Integration Status from Low-Grade Cervical Lesion to Invasive Cervical Cancer

Cheung¹,

Cheung²,

Ng³

et al. 2009

J Clin Microbiol

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The clinical value of viral load and integration testing for human papillomavirus (HPV) remains unclear. Data on HPV type 18 (HPV18) is limited. We examined the HPV18 viral load and integration status of 78 women with normal cervix or neoplasia. While the crude viral load appeared to increase with lesion severity, the association was not significant after normalization with sample cellularity. Unlike reports for HPV16, the amino-terminal 1 region of HPV18 E2 was most frequently (71.0%) disrupted, representing the best marker for integration. A substantial proportion (57.1%) of invasive cancers harbored only the episomal genome, thus jeopardizing the clinical value of integration testing. A large proportion (41.7%) of normal/low-grade lesions showed viral integration, suggesting that integration of HPV18 starts early and is unlikely to be a sole determinant for progression. Interpretation of viral load should take into account the form of HPV infection as single infections had significantly higher viral loads than coinfections (P ‫؍‬ 0.046). More data generated from routinely collected samples are warranted to verify the clinical value of viral load and integration testing. Viral load quantitation for HPV18 is premature for clinical use at this stage.There is no doubt that infection with human papillomavirus (HPV) may lead to the establishment of cervical cancer (3, 33). Though infection with HPV is common, not all HPV types will lead to cancer. There are 15 high-risk HPV types that have a higher propensity for the development of cervical cancer, with HPV type 16 (HPV16) and HPV18 being the most prevalent high-risk types worldwide (8,25).Women who have tested positive for high-risk HPV tend to harbor abnormal cervical cytology, and some studies have reported a correlation between viral load and disease severity (19,20,24,28,31). However, the consistency of these findings has been questioned, and it is now realized that the relationship is a lot more complex (5-7, 35). As the severity of disease increases, HPV is often found integrated into the host genome, making the interpretation of viral load even more complex. Furthermore, the available viral load data have been generated using different methodologies that may not be directly comparable (13,17,28,29).The HPV genome can exist in two physical forms: a closed circular episomal form or linearized and integrated into the host genome. Integration of the viral genome is often associated with the disruption of the E2 gene, which has regulatory control of the oncogenes E6 and E7. Nevertheless, this concept has been questioned by a recent study where the integration of the HPV16 genome was not necessarily associated with the overexpression of E6 and E7 (14). HPV18 has been found to be strongly associated with adenocarcinoma of the cervix. Studies carried out with HPV18 have shown that the viral genome is almost invariably found to be integrated in highgrade cervical intraepithelial lesions and invasive cancers (2,9,22,27,32), and these findings potentially allow the us...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analysis of Human Papillomavirus Type 18 Load and Integration Status from Low-Grade Cervical Lesion to Invasive Cervical Cancer

Cheung¹,

Cheung²,

Ng³

et al. 2009

J Clin Microbiol

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“…Its composition (chimeric preparation of HPV18 and human albumin targets) was designed such that it would not interfere with any of the organisms found in stool specimens. A chimeric nucleotide fragment was constructed as previously described for quantification of the IS6110 gene copy number (Carcopino et al, 2006;Menard et al, 2008) (Fig. 2).…”

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confidence: 99%

Detection of Mycobacterium tuberculosis complex organisms in the stools of patients with pulmonary tuberculosis

et al. 2009

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The laboratory diagnosis of pulmonary tuberculosis mainly relies on the detection of Mycobacterium tuberculosis complex (MTC) organisms in the sputum. In patients who do not give sputum, alternative respiratory tract specimens can be obtained only by invasive procedures. Based on the known survival of MTC organisms in the gastric fluid, we hypothesized that swallowed MTC organisms would be detectable in stool samples. We compared the presence of MTC organisms in respiratory tract specimens and stool specimens collected in parallel from the same patients. MTC was detected in cultures grown on egg-based medium after appropriate decontamination, by microscopic examination after Ziehl-Neelsen staining and by real-time PCR detection of IS6110 using internal controls. A case of pulmonary tuberculosis was defined by the presence of (i) clinical and radiological signs and symptoms suggestive of pulmonary tuberculosis, and (ii) culture of MTC organisms from at least one respiratory tract specimen or (iii) the presence of acid-fast bacilli in the sputum that were subsequently identified as MTC organisms by real-time PCR. The observation of 134 patients suspected to be suffering pulmonary tuberculosis led to the identification of 24 cases and 110 non-infected control patients. Cases and controls did not significantly differ with respect to sex but cases were significantly younger than controls. The sensitivity/specificity was 37.5 %/100 % for the microscopic examination of stools, 54.2 %/100 % for culturing and 100 %/97.3 % for real-time PCR. The positive predicted value was 100 %, 100 % and 88.9 %, respectively, and the negative predicted value was 88 %, 90.9 % and 100 %, respectively. In four patients, a stool specimen initially yielded the correct diagnosis of pulmonary tuberculosis before evaluation of the respiratory tract specimen confirmed the diagnosis. These data indicate that stools could be used in conjunction with sputum testing or as an alternative specimen upon which to base the diagnosis of pulmonary tuberculosis by molecular identification of acid-fast bacilli and culture. This non-invasive alternative procedure is of particular interest for patients who cannot expectorate. INTRODUCTIONPulmonary infection is the most prevalent form of tuberculosis (Saranchuk et al., 2007). Exposure to contact persons can cause secondary infection, and in some areas sustained high-level prevalence of this deadly infection (Johansen et al., 2002). The diagnosis of pulmonary tuberculosis currently relies upon the detection of Mycobacterium tuberculosis complex (MTC) organisms in sputum (Pfyffer, 2007). Some patients, however, are unable to produce sputum, including children (Oberhelman et al., 2006;Starke & Correa, 1995), immunocompromised patients and patients with neurological impairment (Andresen, 2007). In these situations, alternative specimens obtained by invasive procedures include nasopharyngeal aspirates (Owens et al., 2007), gastric aspirates (Chierakul et al., 2003), respiratory tract secretions obtained by bronchos...

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“…El análisis de la carga viral se realizó mediante PCR cuantitativa, se observó un amplio rango de valores en LIEBG y LIEAG, inclusive en el grupo sin lesión se observaron valores altos en ocho casos, estos valores altos en mujeres sin lesión son probablemente el reflejo de una infección productiva con mayor riesgo de progresar a LEIBG, dada la asociación observada en este y otros estudios (Carcopino et al, 2006;Flores et al, 2006;Cricca et al, 2007;Saunier et al, 2008;Shukla et al, 2014) entre mayor valor de carga viral y progresión de la enfermedad. Al comparar las medias de carga viral en cada grupo histológico se observó una diferencia estadísticamente significativa, con un incremento a mayor severidad de la enfermedad.…”

Section: Discussionunclassified

“…Al comparar las medias de carga viral en cada grupo histológico se observó una diferencia estadísticamente significativa, con un incremento a mayor severidad de la enfermedad. Los estudios en los que se ha empleado la PCR cuantitativa para evaluar la carga viral han mostrado de manera consistente que a mayor grado de la lesión es significativamente mayor la carga viral (Carcopino et al, 2006;Flores et al, 2006;Cricca et al, 2007;Saunier et al, 2008;Shukla et al, 2014), aunque en algunos estudios no se ha encontrado esta diferencia (Andersson et al, 2005;Briolat et al, 2007;Boulet et al, 2009). Cabe señalar que una comparación directa entre nuestros resultados y los obtenidos previamente por otros grupos es difícil dadas las diferencias en las técnicas de tiempo real empleadas (Taqman, SYBR Green), y las múltiples formas de presentación de los resultados (copias/ célula, copias/ng de ADN, copias/muestra).…”

Section: Discussionunclassified

Integración, carga viral y niveles de ARN mensajero de E2 de VPH 16 en la progresión de lesiones intraepiteliales cervicales

Trujillo¹,

Sánchez²,

Bravo³

2018

Acta biol. Colomb.

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RESUMENEntre las lesiones intraepiteliales escamosas cervicales (LIE) es importante distinguir aquellas asociadas con mayor riesgo de cáncer de cuello uterino. El objetivo de este trabajo fue evaluar si los niveles de expresión de E2 del VPH16 en mujeres con LIE y con evidencia de integración viral se asocian con el grado de la lesión. Se analizaron 109 cepillados cervicales positivos para VPH 16 provenientes de 19 mujeres sin LIE, 45 mujeres con LIE de bajo grado (LIEBG) y 45 mujeres con LIE de alto grado (LIEAG). Se cuantificó el número de copias de ARNm de E2 y de los genes E2 y E6 mediante PCR en tiempo real para determinar la carga viral (E6) y la proporción E2/E6 para evaluar la integración viral. Se encontraron frecuencias similares de expresión de E2 en LEIBG y LEIAG 15/45 (33 %), la frecuencia en mujeres sin lesión fue menor 3/19 (15,8 %), todos los casos en los que se observó expresión del gen E2 tenían mezcla de ADN viral episomal e integrado. La carga viral aumentó significativamente a mayor grado de la lesión (p=0,049), mientras que la proporción E2/E6 disminuyó (p=0,049). El análisis ROC mostró una baja capacidad de los tres parámetros virales para distinguir entre lesiones de bajo y alto grado. En conclusión, aunque las lesiones con presencia de ADN viral mixto e integrado y expresión de E2 podrían estar en menor riesgo de progresión, y la carga viral y la integración se relacionaron con mayor gravedad de la lesión, su valor clínico como biomarcadores de LEIAG es limitado.Palabras clave: carga viral, integración viral, lesión intraepitelial escamosa de alto grado, lesión intraepitelial escamosa de bajo grado, reacción en cadena de la polimerasa en tiempo real, virus del Papiloma Humano tipo 16. ABSTRACTIt is important to distinguish among squamous intraepithelial lesions (SIL) those associated with increased risk cervical cancer. Our aim was to evaluate if the expression level of gen E2 in women with SIL and evidence of viral integration is associated to the grade of lesion. Cervical scrapes HPV16 positive from 19 women with normal histology, 45 women with low-grade SIL (LSIL) and 45 women with high-grade SIL (HSIL) were analyzed. Real-time PCR was used to quantify the mRNA of E2 and E2 and E6 genes to calculate viral load (E6) and the ratio E2/E6 to assess viral integration. Similar frequencies of E2 expression were found in LSIL and HSIL15/45 (33 %), the frequency in women without SIL was lower 3/19 (15.8 %), and all cases with E2 gene expression had mixed episomal and integrated viral DNA. The viral load increased significantly with the grade of SIL (p= 0.049), while E2/E6 ratio decreased (p=0.049). The ROC analysis showed low capacity of the three viral parameters analyzed to distinguish between low and high grade SIL. In conclusion although SIL with mixed and integrated viral DNA with E2 expression could be at lower risk of progression, and viral load and integration were associated with higher severity of the lesion, its clinical value as biomarkers of HSIL is limited.

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Determination of HPV type 16 and 18 viral load in cervical smears of women referred to colposcopy

Cited by 62 publications

References 34 publications

Analysis of Human Papillomavirus Type 18 Load and Integration Status from Low-Grade Cervical Lesion to Invasive Cervical Cancer

Analysis of Human Papillomavirus Type 18 Load and Integration Status from Low-Grade Cervical Lesion to Invasive Cervical Cancer

Detection of Mycobacterium tuberculosis complex organisms in the stools of patients with pulmonary tuberculosis

Integración, carga viral y niveles de ARN mensajero de E2 de VPH 16 en la progresión de lesiones intraepiteliales cervicales

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