Quantification of HIV-1 proviral DNA from peripheral blood mononuclear cells using a high throughput four-competitor competitive PCR.

A multiple competitor PCR (mcPCR) was developed to quantify HIV-1 proviral DNA from peripheral blood mononuclear cells (PBMC). DNA extracted from a mixture of HIV infected PBMC and four size-mutated DNA competitors were co-amplified. The Cy5-fluorescence labelled PCR products were denatured by heating, separated using an automated DNA sequencer and quantified by a fragment analysis computer software. An internal standard was generated by plotting the peak areas of the four competitors against their inputs. Based on the internal standard, HIV sample DNA was quantified by extrapolating the corresponding signal. The linear range of the mcPCR was three log wide and the quantitation limit was about 20 copies of HIV DNA/10(6) PBMC. Using the mcPCR, HIV DNA was quantified from 14 long-term non progressors (LTNP) and 14 patients with advanced disease. A significantly lower copy number of HIV DNA was obtained in the LTNP (p = 0.018). These data suggest that the mcPCR is sensitive, reliable and especially useful for HIV DNA quantification of a large number of clinical samples.

Fatal sclerosing peritonitis associated with primary effusion lymphoma after liver transplantation: a case report.

Sclerosing peritonitis (SP) after liver transplantation has been described in 10 cases in the literature. The etiology is still unknown; however, SP is considered a consequence of chronic irritation and inflammation. It can be classified as primary (idiopathic) or secondary form. Although pathologically benign, it has a negative course, resulting in unrelenting abdominal pain, small bowel obstruction, malnutrition, and death. Posttransplantation lymphoproliferative disease (PTLD) is one of the leading causes of late death. Its development is related to complex interactions between immunosuppressive drugs and environmental agents. Primary effusion lymphoma (PEL) as an onset presentation of PTLD is relatively uncommon. Most examples of effusion-based PTLD have been secondary to widespread solid organ involvement and associated with Human herpes virus 8 (HHV-8) recurrence. Here in, we report a case of a 55-year-old man who rapidly developed refractory ascites and bacterial peritonitis at 1-year after orthotopic liver transplantation (OLT) with a fatal clinical course at the beginning of the second follow-up year after an uncomplicated liver transplantation due to cryptogenic cirrhosis. The diagnosis of HHV-8-positive lymphoma was established by postmortem examination with multiple solid localizations and massive dense fibrotic adhesions encompassing the small intestine, colon, liver, and porta hepatis without any involvement of body cavities.

Chlamydia pneumoniae respiratory infections among patients infected with the human immunodeficiency virus.

Thirteen cases of Chlamydia pneumoniae infection in patients seropositive for the human immunodeficiency virus (HIV) are described. The occurrence, the clinical spectrum, and the significance of the infection during HIV disease are compared with data reported in the literature. Chlamydia pneumoniae infection was established by a serologic micro-immunofluorescence test using standard diagnostic criteria. In four cases the results of serological tests were confirmed by direct immunofluorescence on respiratory specimens. Five patients developed focal pneumonia but recovered completely after specific antibiotic treatment. Three patients developed severe and diffuse interstitial pulmonary involvement, two of whom died of acute respiratory failure. Five patients developed upper respiratory tract infection. Using 39 pair-matched HIV-seropositive subjects as controls, the cases of infection were found to be significantly associated with a previously diagnosed pulmonary disease. Upon retrospective analysis of 319 consecutive cases of pneumonia among HIV-infected patients, Chlamydia pneumoniae was the sole agent detected in eight (2.5%) cases, and Chlamydia pneumoniae together with other infectious agents was detected in seven (2.2%) cases. Chlamydia pneumoniae is a possible cause of severe respiratory infection in Italian HIV-infected immunocompromised patients, and its presence must be suspected when patients do not respond to therapy with beta-lactam agents or to anti-Pneumocystis carinii treatment.

Sporadic hepatitis C virus infection: a case-control study of transmission routes in a selected hospital sample of the general population in Italy.

A case-control study was performed on 9,175 Italian adult outpatients in 5 hospitals in Rome. The study was carried out to clarify the role of some less investigated risk factors (RF) in the spread of hepatitis C virus (HCV) infection. All subjects were contacted by interviewers, who completed a questionnaire. Their sera were stored and subsequently tested for both HCV and hepatitis B virus core (HBc) antibodies. 365 subjects, positive for anti-HCV and anti-HBc-negative, and who had denied intravenous drug use (IDU) (cases) were compared with an equal number of suitable random controls negative for anti-HCV and anti-HBc. Gender, age and region of birth and residence were matched. The prevalence of 13 RFs were statistically compared by univariate and multivariate analysis. A positive anti-HCV test was significantly associated, by multivariate analysis with intravenous treatments and minor surgical procedures (both before 1975) (p < 0.001), blood transfusions (before 1991) (p < 0.01), diabetes (p < 0.01), and deliveries in hospital (p < 0.05) (both before 1975). After 1975 (1991 for transfusions), all associations lost their significance. Intra-familial (sexual and non sexual), occupational RFs and dental care were not significantly associated with the presence of anti-HCV. We suggest that non-disposable syringes, commonly used until 1975 in Italy for i.v. treatments, have been the major route for HCV transmission in Italy among non-IDU subjects.