Human cytomegalovirus infection during childhood: detection of viral DNA in peripheral blood by means of polymerase chain reaction

Shibata, Mitsue; Morishima, Tsuneo; Terashima, Makoto; Kimura, Hiroshi; Kuzushima, Kiyotaka; Hanada, Naoki; Nishikawa, Kiyonobu; Watanabe, Kazuyoshi

doi:10.1007/bf00192462

Cited by 18 publications

(7 citation statements)

References 29 publications

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“…195, 4 ml of 5 mM each dNTPs, 4 ml of 5x reverse transcriptase sackie viruses and Echovirus), with the exception of echovirus 22 and 23 (46). A second primer pair, buffer (51 buffer Å 15 mM MgC12, 50 mM dithiothreitol, 250 mM Tris, pH 8.3, and 375 mM KCl), 200 units designed to amplify a 607-bp product was created from the immediate early gene of the AD169 strain of Maloney murine leukemia virus (MoMLV) reverse transcriptase (BRL-Gibco), and DEPC-treated dH 2 O to of CMV, recognizing all CMV serotypes (33). The third primer pair was designed to amplify a 308-20 ml; this reaction was incubated at 37ЊC for 1 h. Two microliters of this first-strand cDNA or 500 ng of control bp sequence of the hexon region of all adenovirus serotypes (except adenovirus 40,41) (29).…”

Section: Nucleic Acid Extractionmentioning

confidence: 99%

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Detection of Intrauterine Viral Infection Using the Polymerase Chain Reaction

Veyver

Bowles

et al. 1998

Molecular Genetics and Metabolism

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Section: Nucleic Acid Extractionmentioning

confidence: 99%

“…nucleic acid extraction. PCR and RT-PCR were per-Control Samples formed as described previously (15,29,30,33,42,43,47,49,50). Amniotic fluid samples were collected for genetic…”

mentioning

confidence: 99%

Detection of Intrauterine Viral Infection Using the Polymerase Chain Reaction

Veyver

Bowles

et al. 1998

Molecular Genetics and Metabolism

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“…In some studies, HCMV DNA has been detected in the peripheral blood mononuclear cells of healthy blood donors,' 5*28 but other studies have found HCMV DNA only in leukocytes from severely immunosuppressed patients. [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29] In our study, HCMV DNA was detected by amplification in the peripheral blood mononuclear cells of normal healthy individuals who were negative for urinary excretion (data not shown). This is why we used urinary excretion as the method for monitoring HCMV infection.…”

Section: Discussionmentioning

confidence: 84%

Increased urinary excretion of human cytomegalovirus in children with malignancy: Detection by polymerase chain reaction

Kajiwara

Yamaguchi

Hirai

et al. 1993

Pediatrics International

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Human cytomegalovirus (HCMV) is one of the most important agents causing opportunistic infections in immunocompromised hosts. In this study, we examined the urinary excretion of HCMV in children with malignancy using polymerase chain reaction (PCR).Urine samples were collected from on-therapy, off-therapy patients with malignancy, and healthy controls. A simple DNA extraction method using glass powder was employed, and inhibitory effect of urine on PCR was prevented. For PCR, a pair of primers from the HCMV major immediate early gene sequence was used.Among patients who received intensive chemotherapy, 52.0% had urinary HCMV excretion after the chemotherapy course. In contrast, off-therapy patients and healthy controls showed a lower incidence of urinary HCMV excretion (20.4 and 8.7%, respectively). The incidence of HCMV urinary excretion in the on-therapy group was significantly higher than healthy controls ( P < 0.05). In the on-therapy group, the total white blood cell count of the virus excreters was lower than that of non-excreters.The incidence of HCMV excretion was high in on-therapy patients. Most of the virus excreters were seropositive, so their viruria was thought to be caused by reactivation. Repeated monitoring of virus excretion by this rapid and simple method may be useful to detect HCMV infection early and to control it in such patients. Abstract Key wordsHCMV, PCR, pediatric cancer patients, reduction of inhibitory effect, urine.

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“…The second group consisted of 20 children with acute or chronic hepatitis of unknown etiology (mean age 2.9 years). Hepatitis A, B, C, and GB virus type C, and infantile hepatitis with herpesviruses (cytomegalovirus or Epstein-Barr virus) were ruled out in these patients [12,22,23,32]. The third group included 18 children who were transfusion recipients or hemophiliacs (mean age 8.2 years).…”

Section: Study Populationmentioning

confidence: 99%

The prevalence of TT virus (TTV) infection and its relationship to hepatitis in children

Iriyama

Kimura

Nishikawa

et al. 1999

Medical Microbiology and Immunology

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TT virus (TTV) is a newly discovered virus from a patient with post-transfusion hepatitis. We investigated the frequency and pathogenesis of TTV infection in children. A semi-nested PCR assay was used to amplify TTV-DNA in serum samples from 254 ambulatory children without liver disease, 20 with hepatitis of unknown etiology, and 18 transfusion recipients or hemophiliacs. In positive samples, TTV-DNA was quantified by real-time quantitative PCR using a fluorescent probe. We detected TTV-DNA in 20% of children with hepatitis of unknown etiology, which was not statistically different from the 23% prevalence in ambulatory children. In transfusion recipients or hemophiliacs, the frequency was higher (50%) than that in ambulatory children (P = 0.01). Among ambulatory children, TTV-DNA was frequently detected in children with acute gastroenteritis (36%). TTV-DNA was detected in 10% of the infants under 6 months old, and 20% of the children from 7 to 12 months old. The prevalence was constant after the age of 1 year; however, the copy number of TTV-DNA was significantly higher in children under 1 year of age (mean: 10(5.4) versus 10(3.8) copies/ml, P= 0.008). Finally, TTV-DNA was quantified serially in three children with chronic hepatitis who were positive for TTV-DNA. The presence or amount of TTV-DNA was unrelated to the serum alanine aminotransferase level. These results indicate that TTV infection is common in children. The larger quantity of TTV-DNA in infants and the high prevalence of TTV in children of all ages suggest that TTV may be transmitted in early childhood. Its relationship to hepatitis is doubtful in children.

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Human cytomegalovirus infection during childhood: detection of viral DNA in peripheral blood by means of polymerase chain reaction

Cited by 18 publications

References 29 publications

Detection of Intrauterine Viral Infection Using the Polymerase Chain Reaction

Detection of Intrauterine Viral Infection Using the Polymerase Chain Reaction

Increased urinary excretion of human cytomegalovirus in children with malignancy: Detection by polymerase chain reaction

The prevalence of TT virus (TTV) infection and its relationship to hepatitis in children

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