AIMTo investigate whether urine is a good medium for screening and whether there is a correlation between the amount of extracted DNA and human papillomavirus (HPV)-positivity.

METHODSIn the present study, 30 first-voided urine (FVU) specimens and 20 urethroglandular swabs using cervex-brushes from male partners of HPV-positive patients, and 31 FVU specimens and 100 liquid-based cervix cytology leftovers sampled with cervix-brushes from HPV-positive women were examined for the presence of beta-globin. Oncogenic HPV were detected using type-specific PCR.

RESULTSbeta-globin was found in all the brushed samples, whereas it was found in only 68.9% of the FVU specimens. HPV-PCR was positive in 60.0% of the male brushes, in 29% of the female brushes and in 0% of the male FVU specimens. DNA concentration was, respectively, 0.9998 ng/microL, 37.0598 ng/microL and 0.0207 ng/microL.

CONCLUSIONUrine is not a good tool for HPV detection, probably because the low DNA concentration reflects a low amount of collected cells. beta-globin is measurable in FVU by real time quantitative PCR, but the DNA concentration is lower compared to brush sampling for both genders. beta-globin-positivity of urethral and cervical swabs is 100%, showing a higher mean concentration of DNA, leading to a higher detection rate of HPV. This is the first article linking DNA-concentration to the presence of HPV.

Alphapapillomavirus ; genetics ; isolation & purification ; Cervix Uteri ; virology ; DNA, Viral ; genetics ; isolation & purification ; Female ; Globins ; urine ; Humans ; Male ; Papillomavirus Infections ; diagnosis ; epidemiology ; urine ; Polymerase Chain Reaction ; Urine ; virology

OBJECTIVETo quantify human cytomegalovirus (HCMV) DNA in the blood and urine of infants of different ages with suspected HCMV infection, and in the breast milk of their mothers, and to evaluate the significance of HCMV DNA detection in the three specimen types in the diagnosis of HCMV infection among infants of different ages.

METHODSA total of 170 infants with suspected HCMV infection were divided into groups A (<28 days; n=43) and B (28 days to 5 months; n=127) according to their ages. Blood and urine were collected from the infants, and breast milk was collected from their mothers. The specimens were examined by fluorescence quantitative PCR for detection of HCMV DNA.

RESULTSIn group A, HCMV DNA detection rates in blood, urine and breast milk were 65.1%, 18.6% and 93.0% respectively. In group B, HCMV DNA detection rates in blood, urine and breast milk were 64.6%, 92.9% and 72.4% respectively. HCMV DNA detection rate in urine in group B was significantly higher than in group A (P<0.01), however, HCMV DNA detection rate in mothers' breast milk in group B was significantly lower than in group A (P<0.01). Among the 82 infants with positive results for blood and urine, the copy number of HCMV DNA in urine was significantly higher than that in blood.

CONCLUSIONSHCMV DNA detection rates in urine and breast milk are different among infants of different ages, so use of suitable specimens according to age is of great significance for improving detection rate.

Animals ; Cytomegalovirus Infections ; diagnosis ; DNA, Viral ; analysis ; blood ; urine ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Milk, Human ; virology ; Pregnancy

OBJECTIVETo investigate the risk factors for hearing impairment induced by cytomegalovirus (CMV) infection in children.

METHODSOne hundred and fifty-eight children diagnosed with CMV infection were enrolled as subjects. Based on the results of the brainstem auditory evoked potential (BAEP) test, patients were classified into normal hearing group (n=117; BAEP≤35) and abnormal hearing group (n=41; BAEP>35). A retrospective analysis was performed on the general information, routine blood indices, liver function, copy number of CMV-DNA in urine and breast milk. The receiver operating characteristic (ROC) curve was used to predict the copy number of CMV-DNA resulting in abnormal BAEP. The Spearman rank correlation analysis was used to test the correlations of the copy number of CMV-DNA in urine with the degree of hearing impairment and platelet count.

RESULTSThe incidence rates of platelet abnormality and abnormal liver function and the copy number of CMV-DNA in urine were significantly higher in the abnormal hearing group than in the normal hearing group (P<0.01). According to the ROC curve, the copy number of CMV-DNA in urine had a sensitivity of 46.3% and a specificity of 93.2% in predicting hearing impairment when it reached 1.415×10(6) per mL. The results of correlation analysis showed that the degree of hearing impairment was positively correlated with the copy number of CMV-DNA (r=0.382, P<0.01); the platelet count was negatively correlated with the copy number of CMV-DNA in urine (r=-0.233, P=0.003).

CONCLUSIONSAn increased copy number of CMV-DNA in urine might be a risk factor for hearing impairment induced by CMV infection. Children are likely to have hearing impairment when the copy number of CMV-DNA reaches 1.415×10(6) per mL. The monitoring of hearing should be strengthened in CMV-infected children with a decreased platelet count.

Cytomegalovirus Infections ; complications ; DNA, Viral ; urine ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Loss ; etiology ; Humans ; Infant ; Infant, Newborn ; Male ; Platelet Count ; ROC Curve ; Retrospective Studies ; Risk Factors

A considerable number of adult Korean women avoid a Pap smear due to fear and discomfort of the pelvic examination. A reliable but noninvasive and comfortable screening method would considerably increase the participation rate. To evaluate the clinical efficacy of urine-based human papillomavirus (HPV) detection by oligonucleotide microarray, the results of HPV test from matched cervical swab specimens were compared. HPV DNA was detected in 70 of 100 cervical samples. HPV 16 was the most prevalent type (38/70), followed by types 18, 58, 52, 33, 35, 31, and 51. HPV DNA was identified in 47 of 90 urine samples. HPV 16 was the most prevalent type (30/45), followed by types 18, 52, 35, 51, 58, 33, and 56. The HPV detection rates of the cervical swabs increased in accordance with the severity of the cytologic and histologic diagnosis. The type specific agreement of HPV DNA tests between cervical swabs and urine was good in HPV 16 (kappa index=0.64 [95% CI: 0.50-0.79]), 18, 52, and 58 and fair in HPV 33 and 35. We propose that a urine HPV test is a valuable adjunctive method for a conventional Pap smear and can be used in population screening for cervical cancer in countries where it is difficult to obtain colposcopic specimens for cultural or religious reasons.
Vaginal Smears ; Uterine Cervical Neoplasms/*diagnosis ; Papillomaviridae/genetics/*isolation & purification ; Oligonucleotide Array Sequence Analysis ; Middle Aged ; Humans ; Female ; DNA, Viral/*urine ; Cervical Intraepithelial Neoplasia/diagnosis ; Aged ; Adult

OBJECTIVTo investigate the potential value of urine hepatitis B virus (HBV) DNA as a new noninvasive diagnostic indicator for HBV-associated glomerulonephritis (HBV-GN).

METHODSA total of 152 patients including 66 with HBV-GN, 66 with non-HBV-GN, and 20 with chronic hepatitis B (CHB) without renal disease were examined for serum and urine HBV DNA levels using polymerase chain reaction (PCR) and for 5 serum HBV markers using enzyme-linked immunosorbent assays.

RESULTSTwenty-two patients (33%) in the HBV-GN group, but none in the other two groups, were found positive for urine HBV DNA. In the diagnosis of HBV-GN, urine HBV DNA had a high specificity (0.98), a good positive predictive value (PPV, 0.96), and a modest negative predictive value (NPV, 0.60). Urine HBV DNA, alone or in combination with serum HBeAg, was superior in the diagnosis of HBV-GN to the combination of urine HBV DNA with serum HBV DNA, hepatitis B surface antigen and the hepatitis B e antigen.

CONCLUSIONUrine HBV DNA may be one of the new noninvasive diagnostic criterion for HBV-GN.

Biomarkers ; blood ; urine ; DNA, Viral ; blood ; urine ; Enzyme-Linked Immunosorbent Assay ; Glomerulonephritis ; diagnosis ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; complications ; Humans ; Polymerase Chain Reaction ; Predictive Value of Tests ; Sensitivity and Specificity

OBJECTIVETo determine the relationship between viral burden in urine and hearing loss in neonates with cytomegalovirus (CMV) infection.

METHODSTwenty-two neonates with CMV infection between April 2006 and January 2010 were enrolled. Their viral burden in urine and hearing loss information were studied. The receiver operating characteristic curve (ROC) was constructed and the cutoff was determined based on their medical information. The hearing levels were evaluated by brain stem auditory evoked potential (BAEP) during the age of 3 to 6 months in 20 patients.

RESULTSThe viral burden in urine in neonates with abnormal BAEP was higher than that in neonates with normal BAEP (5.06 ± 1.50 vs 3.73 ± 0.86, P<0.05). Hearing loss was predicted with a sensitivity of 0.545 and a specificity of 1.0 by using ROC at the cutoff point of 5.1 which were defined after logarithmic conversion at 1.27×10(5) copies/mL of CMV burden in urine. The incidence of hearing loss during the age of 3 to 6 months was strikingly higher in high viral burden group than that in low viral load group (P<0.05).

CONCLUSIONSThe viral burden in urine can predict the possibility of hearing loss in neonates with CMV infection. Hearing loss is likely to be developed when viral burden in urine ≥1.27×10(5) copies/mL in neonates with CMV infection.

Cytomegalovirus ; isolation & purification ; Cytomegalovirus Infections ; complications ; DNA, Viral ; urine ; Evoked Potentials, Auditory, Brain Stem ; Female ; Follow-Up Studies ; Hearing Loss ; etiology ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Viral Load

OBJECTIVETo study the correlation between polyoma virus load and hemorrhagic cystitis after allogeneic stem cells transplantation for prevention of hemorrhagic cystitis.

METHODSBlood and urine specimens were collected from 40 healthy persons, 40 patient with stem cells transplantation and 20 cases complicated with hemorrhagic cystitis for determination of VP1 gene of polyomaviruses BK virus (BKV)/Jamestown Canyon virus (JCV) and simian virus 40 (SV40) by polymerase chain reaction (PCR) and EvaGreen stain fluorescence quantitative assay.

RESULTSIn the peripheral blood, all genes of BKV/JCV and SV40 were negative, while BKV gene in urine and blood from healthy persons and patient with stem cells transplantation was 15% (6/40) and 100% (40/40), respectively. The gene of JCV was positive in 10% (4/40) and 12% (5/40), the gene of SV40 was negative.

CONCLUSIONGenes of BKV and JCV was detectable in urine specimens of healthy persons and there was a correlation between the load of polyomavirus and incidence of hemorrhagic cystitis.

Capsid Proteins ; genetics ; Cystitis ; diagnosis ; etiology ; virology ; DNA, Viral ; blood ; genetics ; urine ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Hemorrhage ; diagnosis ; etiology ; virology ; Humans ; Polymerase Chain Reaction ; methods ; Polyomavirus ; genetics ; growth & development ; Polyomavirus Infections ; complications ; virology ; Viral Load

OBJECTIVETo analyze the relationship between BK viruria and the late onset hemorrhagic cystitis (LOHC) after hematopoietic stem cell transplantation (HSCT), and investigate the role of BK virus load in the development of LOHC.

METHODSFrom August 2006 to April 2008, urine samples were collected weekly from 113 patients undergoing HSCT. Virus DNA were extracted from the urine samples and amplified by qualitative PCR. Real-time quantitative PCR was used to quantify BKV DNA in the urine samples of all BK viruria patients.

RESULTSLOHC occurred in 22 patients (19.5%), including grade 1 in 7, grade 2 in 11, grade 3 in 3, and grade 4 in one. The median onset time was 44 (13 - 114) days after transplantation. Twenty-one of which (95.5%) were BK virus positive, being significantly higher than that in non-LOHC patients (31.9%) (P = 0.000). No BK virus was detected in the healthy control group at the same time. Quantitative PCR detection showed that the mean BK virus DNA copies in LOHC patients at a week before occurring HC was higher than that at the first positive samples (10(5) copies/microl versus 10(4) copies/microl, P = 0.025), and it was no significant change at the onset and a week after HC. Meanwhile, there was no statistical difference in the mean level of BK virus DNA copies among the LOHC patients with different grades. The mean level of BK virus DNA copies in non-HC patients was 10(3) to 10(4) copies/microl, being lower than that in LOHC patients.

CONCLUSIONSBK viruria is an important pathogenic cause of the LOHC after HSCT. The occurrence of BKV viruria in HSCT patients, together with the increasing of BK virus DNA copies in urine, over the level of 10(5) copies/microl may indicate a possible development of LOHC.

Adolescent ; Adult ; BK Virus ; isolation & purification ; Child ; Cystitis ; etiology ; virology ; DNA, Viral ; urine ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Postoperative Complications ; etiology ; virology ; Transplantation, Homologous ; Viral Load ; Young Adult

OBJECTIVETo investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.

METHODFresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared.

RESULTTotally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05).

CONCLUSIONGenotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.

Amino Acid Sequence ; Base Sequence ; Child, Preschool ; Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; DNA Primers ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Pneumonia, Viral ; virology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Urine ; virology ; Viral Envelope Proteins ; genetics

OBJECTIVETo evaluate the detection of cytomegalovirus (CMV) by polymerase chain reaction (PCR) for predicting the development of CMV disease.

METHODSOne hundred and thirty one allo-HSCT patients performed in the past 2 years were analyzed retrospectively. PCR-CMV was used to monitor CMV viremia and vireuria once a week after transplantation.

RESULTSIn the dynamic detection, CMV viremia was positive for at least one chance in 89 patients, vireuria did in 99 patients. Thirty-seven patients developed CMV disease with an accumulative incidence of 32.5%. The incidence of CMV disease was 15.6% in plasma CMV-PCR negative group, 31.3% in positive once group, and 47.3% in positive over twice group. There was significant difference among the three groups (P = 0.0126). The incidence of CMV disease was 24.8% in urine CMV-PCR negative group, 43.5% in positive once group, and 33.0% in positive over twice group, being no significant difference among them (P = 0.845). On analysis, viremia could predict the development of CMV disease: the PPV (positive predictive value) is 40.5%, NPV (negative predictive value) is 84.4%, sensitivity is 75.0%, and specificity is 69.2%.

CONCLUSIONSDetected by CMV-PCR, MCV viremia may predict the development of CMV disease, but MCV vireuria cannot.

Adolescent ; Adult ; Child ; Child, Preschool ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; diagnosis ; etiology ; DNA, Viral ; blood ; urine ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Sensitivity and Specificity ; Transplantation, Homologous ; adverse effects