Evaluation of a rapid immunochromatographic test for the detection of norovirus in stool samples.

Worldwide noroviruses are an important cause of gastroenteritis and are major agents of both sporadic as well as epidemic infection. Because of the rapid transmission of the virus, early detection is essential. Until recently, the available test methods for the detection in stool were enzyme immunoassays and real-time reverse transcription polymerase chain reaction (RT-PCR), both of which take several hours to perform. We evaluated the rapid immunochromatographic test RIDA(R)QUICK Norovirus for the detection of norovirus in the stool of patients with acute gastroenteritis. This test is easy to perform and read and only takes 20 min. The sensitivity and specificity compared to RT-PCR results and the positive and negative predictive values were 57.1%, 99.1%, 93.3% and 91.2%, respectively. The rapid test is useful for quick screening, but a negative result should be followed up by RT-PCR.

Monitoring the potential introduction of the Swedish Chlamydia trachomatis variant (swCT) in the Netherlands.

This report describes the actions of public health experts in cooperation with specialists in sexually transmitted diseases (STD), epidemiologists and (molecular) microbiologists to investigate the possible introduction of the swCT variant in the Netherlands: 1. Investigating trends in CT epidemiology Result: STD surveillance and laboratory surveillance did not show any evidence of the introduction of the swCT variant in Holland. 2. Retesting samples by TaqMan PCR Result: Roche CT-negative samples suspected to be CT-positive on the basis of the clinical picture were retested by swCT TaqMan but did not harbour the swCT variant 3. Screening sample pools for the presence of the swCT variant Result: Four different sample pools covering a wide geographical range were tested by specific swCT Taqman assay, but the swCT variant was not detected in any of them. In conclusion, to date the swCT variant has not been found in the Netherlands. However, ongoing monitoring is needed until Roche and Abbott have adapted their CT nucleic acid amplification tests (NAATs) to detect the new variant.

Estradiol-induced synthesis of vitellogenin. I. The isolation of large polysomes from estrogenized rooster liver.

Conventional procedures for the isolation of polysomes, applied to estrogenized rooster liver, fail to yield polysomes containing 30 or more ribosomes, the size expected for polysomes synthesizing the estradiol-induced protein vitellogenin. A new procedure characterized by early and selective removal of cell components ribonucleases allowed the isolation of polysomes with up to 55 ribosomes. Electron microscopy was used for the determination of polysome size and showed that the large polysomes were not aggregates.

Prediction of recurrent cytomegalovirus disease after treatment with ganciclovir in solid-organ transplant recipients.

CMV disease often recurs after initially successful antiviral therapy. We retrospectively determined in a group of 36 organ transplant patients whether clinical, virological, or immunological parameters during or shortly after cessation of antiviral therapy can identify those at high risk of relapse. Eleven of 36 patients had recurrent CMV disease after ganciclovir therapy. Neither donor or recipient CMV serostatus, type of baseline immunosuppression, antirejection treatment, indication for antiviral treatment, nor presence of CMV in the blood during or after therapy (as detected by antigenemia, viremia, or a positive polymerase-chain-reaction signal) were helpful in identification of patients with subsequent relapse. However, quantitative monitoring of antigenemia fascilitated early diagnosis of relapse since 10 of 11 patients with > or = 10 antigen-positive cells per 50,000 PMNs relapsed (99.1%, 95% CI 58.7-99.8). IgM and IgG responses against CMV during primary infection were comparable in relapsing and nonrelapsing patients. During secondary infection relapse occurred only in the 4 patients with the lowest IgG responses. The number of activated CD8bright lymphocytes in the peripheral blood as determined by flow cytometry at the end of antiviral therapy was a strong risk factor for the subsequent clinical course: 6 of 7 patients (85.7%, 95% CI 42.1-99.6%) with < 100 x 10(3) HLADR+CD8bright cells/ml blood relapsed, while 8 of 8 (100%, 95% CI 63-100) with activated CD8bright cells above that level remained asymptomatic (P < .025). These data show that patients with a high risk of relapse of CMV disease can be identified at the end of antiviral therapy.

Cytomegalovirus antigenemia as a useful marker of symptomatic cytomegalovirus infection after renal transplantation--a report of 130 consecutive patients.

In earlier work we demonstrated that CMV immediate early antigens can be detected in peripheral blood leukocytes of patients with active CMV infection. We now report a comparison of the antigenemia assay and an anti-CMV ELISA in a prospective longitudinal study of 130 renal transplant recipients who were monitored for active CMV infection during the first 3 months after transplantation. Active CMV infection developed in 56 patients. The antigenemia assay had a sensitivity of 89% and a specificity of 93% in the diagnosis of active CMV infection; for the ELISA these figures were 95 and 100%, respectively. In 22 of the 56 patients a CMV syndrome occurred. Antigenemia was demonstrated in all 22 patients while an antibody response occurred in 21 of them. The antigenemia assay became positive 8 +/- 7 days before the onset of symptoms while the antibody response was observed 4 +/- 9 days after the onset of symptoms. The pattern of antigenemia was helpful for monitoring the course of the infection. The maximum level of antigenemia was significantly higher and its duration significantly longer in symptomatic than asymptomatic infection. We conclude that CMV antigenemia is a sensitive, specific, and early marker of CMV infection. The antigenemia assay is of great value in monitoring patients with a high risk of CMV infection.

Bell's palsy and herpes simplex virus.

Bell's palsy, which is defined as idiopathic peripheral facial paralysis of sudden onset, accounts for > 50% of all cases of facial paralysis. Different theories on the etiology of Bell's palsy have been proposed and investigated. Various clinical studies have suggested an etiological link between Bell's palsy and herpes simplex virus (HSV). In addition, animal experiments have shown the ability of HSV to induce facial paralysis. In our opinion, the possible link between Bell's palsy and HSV can only be explored properly by studying the human facial nerve, and especially the geniculate ganglion itself. Different groups have tried to detect hypothetically reactivated and hypothetically latent HSV in the facial nerves of Bell's palsy patients and control patients, respectively. The isolation of infectious HSV from facial nerve tissue by conventional cell culture methods appeared to be very difficult, also when Bell's palsy patients were tested. Instead, modern molecular methods, such as in situ hybridization and the polymerase chain reaction (PCR) could easily detect HSV DNA in geniculate ganglia. The detection of HSV-specific latency-associated transcripts in the ganglia of control patients provided further evidence for the hypothetically latent state of HSV in the geniculate ganglia in these patients. Recent PCR experiments performed by a Japanese group strongly suggest that the area adjacent to the geniculate ganglia does not usually contain any HSV at all, except in patients with Bell's palsy. This well-controlled study provides conclusive evidence that reactivation of HSV genomes from the geniculate ganglia is the most important cause of Bell's palsy. Consequently, it has been suggested that "Bell's palsy" be renamed as "herpetic facial paralysis".

Comparison of the Murex Hybrid Capture CMV DNA (v2.0) assay and the pp65 CMV antigenemia test for the detection and quantitation of CMV in blood samples from immunocompromised patients.

BACKGROUND: Presently the semiquantitative pp65 cytomegalovirus (CMV) antigenemia test on white blood cells is often used for monitoring transplant patients for the appearance of active CMV infections. However, the need for immediate processing of the specimens and the lack of interlaboratory standardization of the test may sometimes be a disadvantage. OBJECTIVE: The aim of this study was to investigate the value of the recently developed second version of the Murex Hybrid Capture (MHC) CMV-DNA assay (v 2.0) in comparison with the CMV-pp65 test. The MHC CMV-DNA assay is a quantitative solution hybridization antibody capture assay, using alkaline phosphatase conjugated antibodies and a chemiluminescent substrate. STUDY DESIGN: 248 EDTA blood samples from 33 renal transplant patients and 32 samples from 22 other immunocompromised patients were tested by both the MHC CMV-DNA assay and the CMV-pp65 test. RESULTS: The qualitative ( + or -) results of both tests showed an overall concordance of 81.4%. Calculations on the basis of discordancy analyses showed that the sensitivity, the specificity, and the positive and negative predictive values were 87.7, 98.3, 98.6, 85.2% for the MHC CMV-DNA assay and 76.6, 100, 100, 75.5% for the CMV-pp65 test. Comparison of the quantitative results of both tests systems showed a correlation coefficient of 0.837. In addition, retesting of 50 samples with the MHC-CMV-DNA assay showed an excellent reproducibility with a correlation coefficient of 0.992. All patients which were tested regularly (at least five samples) became either positive with both tests or with none of them. Neither test system detected CMV significantly earlier than the other one. Both tests became strongly positive in all transplant patients with symptomatic CMV infections, and both tests showed a rapid decline of CMV during subsequent antiviral treatment. CONCLUSION: The quantitative Murex Hybrid Capture CMV-DNA assay (v 2.0) may become a valuable additional tool in CMV diagnosis. Further studies will be needed to support this preliminary judgement.

Further details on sequelae at the cervical and tubal level of Chlamydia trachomatis infection in infertile women.

OBJECTIVE: Relationships were studied between correlates of sexually transmitted diseases and chlamydial antibodies versus cervical mucus abnormalities and tubal abnormalities. An estimate was also attempted of the baseline prevalence of endogenous tubal abnormalities. DESIGN: Prospective, descriptive. SETTING: The outpatient department of the Fertility Unit of the Department of Obstetrics and Gynaecology of the Groningen University Hospital. PATIENTS: Females (n = 184) of infertile couples consecutively visiting the department for evaluation and treatment. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Sexually transmitted disease correlates, cervical chlamydial infection, cervical and serum chlamydial antibodies, cervical mucus qualities according to World Health Organization criteria, tubal abnormalities. RESULTS: Of the 175 cervical mucus samples studied, subnormal quality was because of imperfect timing of collection of 22 of the 23 subnormal samples. Tubal abnormalities correlated with more than five lifetime sexual partners (P less than 0.035), as well as with serum immunoglobulin G antibodies (P less than 0.05), which could be because of interaction between both risk factors. CONCLUSIONS: No correlation was found between subnormal cervical mucus and chlamydial infection. Approximately two thirds of the tubal abnormalities seem unrelated to exogenous factors, although further study seems warranted.

Is serology of any use when searching for correlations between Chlamydia trachomatis infection and male infertility?

Evidence on Chlamydia trachomatis causing male infertility is conflicting. We therefore collected data on epidemiological and clinical correlates of chlamydial infection and male fertility in 184 males visiting our Fertility Unit. Antibodies against Chlamydia trachomatis in serum and semen were also determined. Significant correlations were demonstrated between current chlamydial urethral infection and semen immunoglobulin (Ig) A, serum IgA and serum IgG. These parameters, however, were neither related to a history of sexually transmitted disease nor to lifetime number of sexual partners. Why, in the male, serology does not correlate with chlamydial infection in a more remote past is explained. Our data support, on epidemiological as well as serological grounds, the conclusion that chlamydial infection probably does not contribute significantly to male infertility.

Direct immunofluorescence for Chlamydia trachomatis on urogenital smears for epidemiological purposes.

A population of 197 asymptomatic women, attending an out-patient department for birth-control advice, was screened for urogenital infection with Chlamydia trachomatis by direct immunofluorescence on cervical and urethral smears. A blood sample was obtained for chlamydial serology and demographic, behavioural and clinical data were recorded. Eleven (5.5%) women had a chlamydial infection. Chlamydial infection, as diagnosed with direct immunofluorescence, correlated with a history of sexually transmitted disease (p less than 0.01), promiscuity (p less than 0.01), use of oral contraceptives (p less than 0.02) and high chlamydial antibody titres (p less than 0.01). These last also correlated with a history of sexually transmitted disease (p less than 0.02) and promiscuity (p less than 0.02). These results, obtained with direct immunofluorescence, are indistinguishable from those obtained previously with chlamydial culture. Direct immunofluorescence on urogenital smears seems a valuable tool for epidemiological investigation. Our data also support the hypothesis that oral contraceptive use is correlated with chlamydial infection because of increased cervical susceptibility to infection and not because of a sampling bias towards oral contraceptive users.

Epidemiological aspects of chlamydial infections and tubal abnormalities in infertile couples.

Demographic and behavioural data on parameters of previous sexual behaviour were collected of both partners of 184 couples visiting the Department of Obstetrics and Gynaecology because of infertility. They were also screened for urogenital infection with Chlamydia trachomatis. Of 131 (71.2%) women, a diagnosis was established by hysterosalpingography and/or laparoscopy about the condition of their fallopian tubes. An unmarried status correlated significantly with a history of sexually transmitted disease, and with past promiscuity of male and female partner. Abnormalities of the fallopian tubes were significantly related to an unmarried status, a history of pelvic inflammatory disease, and past promiscuity. Male promiscuity was not shown to correlate with tubal abnormalities in the female partner. In only three couples, the partners reported to have had a sexual relationship with a partner other than the current one over the past year. The percentage of Chlamydia trachomatis-positive women of 1.8 and men of 3.0 in our study agrees with this low recent promiscuity. Chlamydia trachomatis was shown to be able to maintain itself for at least 4 years within a couple. The results are commented on.

Therapy of idiopathic sudden sensorineural hearing loss: antiviral treatment of experimental herpes simplex virus infection of the inner ear.

Experimental herpes simplex virus type 1 (HSV-1) labyrinthitis provides a model of idiopathic sudden sensorineural hearing loss (ISSHL). Corticosteroids improve the prognosis for hearing recovery in ISSHL, but the effects of acyclovir are unknown. To establish the therapeutic efficacy of acyclovir (Zovirax) and prednisolone in experimental HSV-1 viral labyrinthitis, we induced HSV-1 labyrinthitis in 12 guinea pigs. Three animals received no treatment, 3 received prednisolone, 3 received acyclovir, and 3 received both. Four other animals served as controls, receiving culture medium only. Hearing, HSV-1 antibody titers, and cochlear damage were evaluated. The HSV-1 labyrinthitis caused hearing loss within 24 hours. Combination treatment consisting of prednisolone and acyclovir resulted in earlier hearing recovery and less extensive cochlear destruction compared to prednisolone or acyclovir as a monotherapy. The beneficial effect of this treatment modality remains to be demonstrated in ISSHL.

[Cause of Bell's palsy: herpes simplex virus]. / Oorzaak van verlamming van Bell: herpes-simplexvirus.

At present Bell's palsy (BP) is still defined as idiopathic unilateral facial paralysis of sudden onset. More than 20 years ago an aetiological link between herpes simplex virus (HSV) and BP was proposed. Numerous experiments in animals and in humans have been performed to test this hypothesis. However, the human facial nerve tissue itself was investigated in only a few of these studies. Two research lines were followed: (a) search for presence of latent HSV in the facial nerve of asymptomatic individuals (as shown earlier in the trigeminal nerve); (b) find 'active' HSV in tissues from BP patients with recent onset of disease. The application of new molecular biological techniques, notably polymerase chain reaction (PCR), to facial nerve tissues has provided ample evidence for a causal relationship between BP and HSV. Therefore it might now be the time to change the name 'Bell's palsy' into 'herpetic facial paralysis'.

[An adenovirus epidemic in patients with kidney transplantation]. / Een adenovirusepidemie onder niertransplantatiepatiënten.

During weekly routine virological screening of kidney transplant patients 12 out of 15 patients within a period of four months were found to be infected with adenovirus. All isolates were of the same serotype, type AdII + 35/HII. However, DNA restriction enzyme analyses showed the presence of two different DNA variants which were associated with three different epidemiological episodes. The epidemic probably started with reactivation of latent virus in a limited number of patients, after which it spread nosocomially. None of the patients showed signs or symptoms clearly attributable to adenoviruses, although adenovirus could not be excluded as a cofactor in the fatal outcome of hepatitis in one of the patients. Adenovirus apparently can easily spread nosocomially. Since literature data suggest that adenovirus infections of transplant patients may result in serious complications, adenovirus should not be neglected in virological screening protocols for kidney transplant patients.

[Chlamydia trachomatis diagnosis in women: the value of serological studies and of culture of cervical and urethral swabs]. / Chlamydia trachomatis-diagnostiek bij de vrouw: de waarde van serologisch onderzoek en van kweek van materiaal uit cervix en urethra.

We studied the value of the determination of the serum IgM antibody level for the diagnosis of genital Chlamydia trachomatis infection in women and the value of the urethra in addition to the cervix as a sampling site for the diagnosis of C. trachomatis infection by culture and direct immune-fluorescence (IF) test. Of 205 women, mean age 26 years (range 14-56), who visited the Sexually Transmitted Disease clinic of the University Hospital Groningen from July 1985 until November 1987, single serum samples were obtained for the determination of IgM and IgG anti-C. trachomatis antibodies using FITC conjugates, in addition to swabs of cervix and urethra for culture and direct IF test. In women with a negative culture and positive IgM serology the culture was repeated. The culture was used as golden standard. In 17/205 (36%) women positive C. trachomatis IgM serology (greater than or equal to 1:32) and in 190/205 (93%) positive IgG serology (greater than or equal to 1:40) was found. In 67/205 (33%) women the C. trachomatis culture of cervix and (or) urethra was positive. The positive and negative predictive values of C. trachomatis IgM serology were 41% and 72%. In 3/20 women with negative culture results and positive IgM serology a second culture was positive. In 176 women, of whom samples from both cervix and urethra were available, the only C. trachomatis positive site by culture was the urethra in 7/54 (13%) and by direct IF in 3/42 (7%). The increase in the prevalence obtained by collecting material not only from the cervix but also from the urethra was 4% by culture and 2% by direct IF.2+n

[Chlamydia trachomatis infection in women and the use of oral contraceptives]. / Chlamydia trachomatis-infectie bij de vrouw en het gebruik van orale anticonceptiva.

We determined the prevalence of genital Chlamydia trachomatis infection in women who visited a clinic for sexually transmitted diseases (STD) and the influence of the number of partners and the use of oral contraceptives (OC), with special attention to the recognition of pelvic inflammatory disease (PID) and to the results of therapy. Of 217 women, with a mean age of 26 years (range 14-56), who visited the STD clinic of the University Hospital of Groningen from July 1985 until November 1987, anamnestic data were collected as well as the results of swabs from cervix and urethra taken for culture and direct immunofluorescence test of C. trachomatis and for gonococcal culture. The influence of the number of partners (1 versus greater than 1) and OC on the prevalence of C. trachomatis infection was evaluated by logistic regression analysis. PID was excluded in coöperation with the department of gynaecology. C. trachomatis-infected women were treated by doxycycline orally (day 1 2 x 100 mg, day 2-7 1 x 100 mg) according to the dosage scheme advised by the Dutch Health Council in 1986. A control culture was taken 2-3 weeks after treatment. C. trachomatis was detected in 72/217 (33%) women by culture and (or) direct IF test and in 22/41 (54%) women with gonorrhoea. In connection with the number of partners in the year preceding the examination, the following prevalences were found: 18/74 (24%) (1 partner), 43/108 (40%) (2-5 partners) and 10/27 (greater than 5 partners).(ABSTRACT TRUNCATED AT 250 WORDS)

PIP: The authors determined the prevalence of genital Chlamydia trachomatis infection in women who visited a clinic for sexually transmitted diseases (STDS) and the influences of the number of partners and of the use of oral contraceptives (OCs), with special attention paid to the recognition of pelvic inflammatory disease (PID) and to the results of therapy. Of 217 women with a mean age of 26 years (range 14-56), who visited the STD clinic of the University Hospital of Groningen from July 1985-November 1987, anamnestic data were collected as well as the results from swabs of the cervix and urethra taken for culture and direct immunofluorescence (IF) test of C. trachomatis and for gonococcal culture. The influence of the number of partners (1 vs 1) and OCs on the prevalence of C. trachomatis infection was evaluated by logistic regression analysis. PID was excluded in cooperation with the department of gynecology. C. trachomatis-infected women were treated by doxycycline orally (day 1, 2x 100 mg, days 2-7, 1x 100 mg) according to the dosage scheme advised by the Dutch Health Council in 1986. A control culture was taken 2-3 weeks after treatment. C. trachomatis was detected in 72/217 (33%) of the women by culture and/or direct IF test and in 22/41 (54%) women with gonorrhea. In connection with the number of partners in the year preceding the examination, the following prevalences were found: 18/74 (24%, 1 partner), 43/108 (40%, 2-5 partners), and 10/27 (5 partners). The prevalence among OC-using women was significantly higher (p0.05) than in non-OC using women: 44.101 (44%) vs 21/93 (23%). This was also true when OC users were compared to sterilized women (9/47, 19%), regardless of number of partners and age. Active PID was found in 2 women, both infected. After treatment with positive C. trachomatis cultures, control cultures were negative in 39/40 (98%). A relatively high prevalence of genital C. trachomatis infection is found in women visiting STD clinics, as well as in women with only 1 partner during the year preceding the examination. This study supports the hypothesis of OC use being a risk factor. How OC use influences PID risk is not fully understood. Within the framework of reliable contraception and prevention of STD complications, the combination of OC + a barrier method should be advised to women without a steady partner. Further study is necessary to determine whether preventive antibiotic treatment of asymptomatic adolescents with anamnestic risk factors for C. trachomatis infection is indicated at the start of a new relationship. (author's modified)

Comparison of two commercial molecular assays for simultaneous detection of respiratory viruses in clinical samples using two automatic electrophoresis detection systems.

Two molecular assays were compared with real-time RT-PCR and viral culture for simultaneous detection of common viruses from respiratory samples: a multiplex ligation-dependant probe amplification (MLPA) and a dual priming oligonucleotide system (DPO). In addition, the positive detections of MLPA and DPO were identified using two different automatic electrophoresis systems. A panel of 168 culture-positive and negative samples was tested by the molecular assays for the presence of influenza A and B virus, respiratory syncytial virus, human metapneumovirus, rhinovirus, coronaviruses, parainfluenza viruses and adenovirus. One hundred and twenty-nine (77%) samples were positive as detected by at least one method. Sixty-nine (41%) samples were positive by cell culture (excluding human metapneumovirus and coronaviruses), 116 (69%) by RT-PCR, 127 (76%) by MLPA and 100 (60%) by DPO. The MLPA yielded results in one attempt for all samples included while 12 (7.2%) samples had to be repeated by the DPO assay due to inconclusive results. The MLPA assay performed well in combination with either electrophoresis system, while the performance of the DPO assay was influenced by the electrophoresis systems. Both molecular assays are comparable with real-time RT-PCR, more sensitive than viral culture and can detect dual infections easily. Results can be obtained within 1 day.