Human papillomavirus detection by PCR assay in a large series of high-grade squamous intraepithelial lesions with cytohistological correlation and follow-up.

OBJECTIVE
High-grade squamous intraepithelial lesions (HSIL) is a precursor of invasive cervical carcinoma and is generally associated with the integration of mucosotropic human papillomavirus (HPV) DNA into the host cell genome. HPV detection is easy to do at this time, even in the laboratory with limited technological capacity, and follow-up procedures for patients with HSIL established.

METHOD
HPV detection is done in a large group of patients with HSIL, and the results were correlated with the findings of cytology, histology, and colposcopy. Differences examined and discussed.

RESULTS
Conventional Papanicolaou (Pap) screening detected 446 HSIL (0.20%) in 218 906 cervical smears. HPV detection by PCR positive in 339/358 (94.7%) patients. Strains involved: HPV 16 in 180 patients (53.1%), HPV 18 in 35 (10.3%), HPV 31/33 in 27 (8%), HPV 6/11 in 10 (2.96%) and of an unknown type in 73 (30%). For the last 97 patients (2006-2007), HPV typing expanded with the following results: HPV 52 was detected in 9 patients (9.2%), HPV 58 in 6 (6.1%), HPV 51 in 4 (4.1 %), HPV 68 in 2 (2.0%), and HPV 39 in 1 (1.0%). The number of patients nonidentified dropped to 9 (9.4%); In addition, 14/97 (14.4%) patients were infected with two or more types of the virus.

Finally, 19 (5.3%) patients with HPV-negative. Colposcopy revealed a small change in 59 patients (17.3%), major changes in 264 (77.6%), and normal findings in 17 (5.1%). A biopsy was taken in 331/446 patients, and the diagnosis of HSIL or overt malignancy was confirmed histologically in 281 (84.9%) patients: in 46 CIN II, CIN III in 224, and histological upgrade in 11 (6 microinvasive squamous carcinoma , 1 squamous carcinoma, 2 endocervical adenocarcinoma in situ, microinvasive endocervical adenocarcinoma and 2). Thirty-five patients (10.6%) relegated to CIN I and 15 (4.5%) patients had negative biopsies. Follow-up in patients with negative biopsies confirmed the existence of SIL in 11 patients [1 HSIL and 10 low-grade squamous intraepithelial lesions (LSIL)] while the fourth is considered a false positive (atrophic changes, 2; changes in reactive, 2).

After treatment, 31/331 (9.36%) patients displayed recurrence (HSIL and LSIL in 29 at 2). Strain of virus involved in patients with relapse were HPV 16 in 16 patients (51.6%); HPV 18 in 4 (12.9%); HPV 16 and 18 in 1 (3.2%); HPV 31 in 1 (3.2%); HPV 52 in 1 (3.2%); HPV 18, 31, and 58 in 1 (3.2%); HPV 68 in 1 (3.2%); HPV 51 and 73 in 1 (3.2%), and of an unknown type in 5 (16.1%). Follow-up in 14/19 HSIL and HPV-negative patients confirmed the existence of cervical pathology.

CONCLUSION
HPV detection improve diagnostic sensitivity and provides an ideal tool for monitoring the response to treatment in patients with HSIL. Pathogenic relevance of HPV strain 18 may be larger than previously thought.

analysis of real-time PCR based on the BRAF V600E mutation in the low and medium grade lymphoma assert common in hairy cell leukemia.

Hairy cell leukemia (HCL) is a rare type of B-cell non-Hodgkin lymphoma (B-NHL), which is not known to be associated with recurrent abnormality karyotype characteristics. A recent study used massively parallel whole exome sequencing to identify mutations in the BRAF V600E enabled, which appears on the HCL.

Here, we confirmed the specificity of BRAF V600E for HCL among low- and middle-grade B-NHL and describes a method of polymerase chain reaction real-time to detect this mutation in cases with a low tumor burden. The V600E mutation does not appear associated with microsatellite instability, unlike the case of colorectal cancer.

MULTIPLEX KIT PCR MASTITIS PCR kit
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Thus, in conjunction with previous data, our results suggest the incorporation of BRAF V600E mutation analysis in the diagnostic workup of cases HCL. In addition, targeting the Ras-Raf-Mek-Erk-Map kinase should be investigated as a potential therapeutic strategy for patients with this disease.