Real-time PCR detection of human herpesvirus 1-5 in patients lacking clinical signs of a viral CNS infection.

BACKGROUND: Infections of the central nervous system (CNS) with herpes- or enterovirus can be self-limiting and benign, but occasionally result in severe and fatal disease. The polymerase chain reaction (PCR) has revolutionized the diagnostics of viral pathogens, and by multiple displacement amplification (MDA) prior to real-time PCR the sensitivity might be further enhanced. The aim of this study was to investigate if herpes- or enterovirus can be detected in cerebrospinal fluid (CSF) from patients without symptoms. METHODS: Cerebrospinal fluid (CSF) samples from 373 patients lacking typical symptoms of viral CNS infection were analysed by real-time PCR targeting herpesviruses or enteroviruses with or without prior MDA. RESULTS: In total, virus was detected in 17 patients (4%). Epstein-Barr virus (EBV) was most commonly detected, in general from patients with other conditions (e.g. infections, cerebral hemorrhage). MDA satisfactorily amplified viral DNA in the absence of human nucleic acids, but showed poor amplification capacity for viral DNA in CSF samples, and did not increase the sensitivity for herpes virus-detection with our methodology. CONCLUSIONS: Viral pathogens are rarely detected in CSF from patients without signs of CNS infection, supporting the view that real-time PCR is a highly specific method to detect symptomatic CNS-infection caused by these viruses. However, EBV may be subclinically reactivated due to other pathological conditions in the CNS.

Extended antimicrobial treatment of bacterial vaginosis combined with human lactobacilli to find the best treatment and minimize the risk of relapses.

BACKGROUND: The primary objective of this study was to investigate if extended antibiotic treatment against bacterial vaginosis (BV) together with adjuvant lactobacilli treatment could cure BV and, furthermore, to investigate factors that could cause relapse. METHODS: In all, 63 consecutive women with bacterial vaginosis diagnosed by Amsel criteria were offered a much more aggressive treatment of BV than used in normal clinical practice with repeated antibiotic treatment with clindamycin and metronidazole together with vaginal gelatine capsules containing different strains of lactobacilli both newly characterised and a commercial one (109 freeze-dried bacteria per capsule). Oral clindamycin treatment was also given to the patient's sexual partner. RESULTS: The cure rate was 74.6% after 6 months. The patients were then followed as long as possible or until a relapse. The cure rate was 65.1% at 12 months and 55.6% after 24 months. There was no significant difference in cure rate depending on which Lactobacillus strains were given to the women or if the women were colonised by lactobacilli. The most striking factor was a new sex partner during the follow up period where the Odds Ratio of having a relapse was 9.3 (2.8-31.2) if the patients had a new sex partner during the observation period. CONCLUSIONS: The study shows that aggressive treatment of the patient with antibiotics combined with specific Lactobacillus strain administration and partner treatment can provide long lasting cure. A striking result of our study is that change of partner is strongly associated with relapse of BV. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01245322.

An outline for a vocabulary of nominal properties and examinations--basic and general concepts and associated terms.

Scientists of disciplines in clinical laboratory sciences have long recognized the need for a common language for efficient and safe request of investigations, reporting of results, and communication of experience and scientific achievements. Widening the scope, most scientific disciplines, not only clinical laboratory sciences, rely to some extent on various nominal examinations, in addition to measurements. The 'International vocabulary of metrology--Basic and general concepts and associated terms' (VIM) is designed for metrology, science of measurement. The aim of the proposed vocabulary is to suggest definitions and explanations of concepts and terms related to nominal properties, i.e., properties that can be compared for identity with other properties of the same kind-of-property, but that have no magnitude.

Clinical laboratory sciences data transmission: the NPU coding system.

In health care services, technology requires that correct information be duly available to professionals, citizens and authorities, worldwide. Thus, clinical laboratory sciences require standardized electronic exchanges for results of laboratory examinations. The NPU (Nomenclature, Properties and Units) coding system provides a terminology for identification of result values (property values). It is structured according to BIPM, ISO, IUPAC and IFCC recommendations. It uses standard terms for established concepts and structured definitions describing: which part of the universe is examined, which component of relevance in that part, which kind-of-property is relevant. Unit and specifications can be added where relevant [System(spec)-Component(spec); kind-of-property(spec) = ? unit]. The English version of this terminology is freely accessible at http://dior.imt.liu.se/cnpu/ and http://www.labterm.dk, directly or through the IFCC and IUPAC websites. It has been nationally used for more than 10 years in Denmark and Sweden and has been translated into 6 other languages. The NPU coding system provides a terminology for dedicated kinds-of-property following the international recommendations. It fits well in the health network and is freely accessible. Clinical laboratory professionals worldwide will find many advantages in using the NPU coding system, notably with regards to an accreditation process.

Changes in the predominant human Lactobacillus flora during in vitro fertilisation.

BACKGROUND: Signature matching of nucleotide sequences in the V1 and V3 regions 16S rRNA genes using pyrosequencing technology is a powerful tool for typing vaginal Lactobacilli to the species level and has been used for investigating the vaginal microbial niche. METHODS: This study has characterized the normal cultivable vaginal Lactobacillus flora at varying estradiol levels in plasma; the study comprised 17 patients undergoing ovarian stimulation for In Vitro Fertilization (IVF) treatment. The vaginal status of each participant was initially assessed as normal according to Amsel and Nugent criteria. RESULTS: L. crispatus, L. gasseri and/or L. jensenii were present in 10 of the patients throughout the study period, and little variation among these three species was encountered in individual patients. The flora of three women was dominated by L. delbrüeckii, L. rhamnosus or L. vaginalis. One woman exhibited a dominance of L. iners. The flora of the remaining three women were initially dominated by L. rhamnosus or L. reuteri, but as their estrogen levels rose, their flora composition altered, to become dominated by one of the three species most common in a normal, healthy vagina. CONCLUSION: Signature matching of nucleotide sequences in the V1 and V3 regions of 16S rRNA genes is a discriminative tool for the study of vaginal Lactobacilli and can be used to track the Lactobacillus flora under a variety of physiological conditions.

Predisposing factors for bacterial vaginosis, treatment efficacy and pregnancy outcome among term deliveries; results from a preterm delivery study.

BACKGROUND: Bacterial vaginosis (BV) during pregnancy is associated with an increased risk of preterm delivery but little is known about factors that could predict BV. We have analyzed if it is possible to identify a category of pregnant women that should be screened for BV, and if BV would alter the pregnancy outcome at term; we have also studied the treatment efficacy of clindamycin. METHODS: Prospective BV screening and treatment study of 9025 women in a geographically defined region in southeast Sweden. BV was defined as a modified Nugent score of 6 and above. Data was collected from the Swedish Medical Birth Register. Women allocated to treatment were supplied with vaginal clindamycin cream. The main outcome goals were to identify factors that could predict BV. RESULTS: Vaginal smears were consistent with BV criteria in 9.3%. Logistic regression indicates a significant correlation between smoking and BV (p < 0.001) and a greater prevalence of BV in the lower age groups (p < 0.001). We found no correlation between BV and history of preterm deliveries, previous miscarriages, extra-uterine pregnancies, infertility problems or reported history of urinary tract infections-factors that earlier have been associated with BV. Treatment with clindamycin cream showed a cure rate of 77%. Less than 1% of women with a normal vaginal smear in early pregnancy will develop BV during the pregnancy. There was no association between BV and the obstetric outcome among women who delivered at term. Women with BV, both treated patients and nontreated, had the same obstetric outcome at term as women with normal vaginal flora. CONCLUSION: BV is more than twice as common among smokers, and there is a higher prevalence in the younger age group. However these two markers for BV do not suffice as a tool for screening, and considering the lack of other risk factors associated with BV, screening of all pregnant women might be a strategy to follow in a program intended to reduce the number of preterm births.

Bacterial vaginosis--a laboratory and clinical diagnostics enigma.

Diagnosing bacterial vaginosis (BV) has long been based on the clinical criteria of Amsel et al., whereby three of four defined criteria must be satisfied. Though there are other criteria and scoring methods which function well in comparison (i.e. Nugent scoring), it is not certain that they will always identify the same category of patients. Point-of-care methods based on various combinations of microbial products, presence of RNA, or more complex laboratory instrumentations such as sensor arrays, have also been introduced for the diagnosis of BV. No method for diagnosing BV can at present be regarded as the best. It could be that--based partly on tacit knowledge on the part of the clinical investigators scoring in the clinic--various scoring systems have been chosen to fit a particular BV-related problem in a particular population. In this review we critically examine these pertinent issues influencing clinical scoring and laboratory diagnostics of BV.

[Can a laboratory investigation be called anything? "The NPU system" sorts out the concepts and gives systematic stringency]. / Får en laboratorieundersökning kallas vad som helst? "NPU-systemet" reder upp i begreppsröran och ger systematisk stringens.

When communicating results from laboratory investigations from the laboratory to the requesters and further between different information systems, it is important that the value as well as the unique identity and name of the laboratory investigation are correctly cited. A committee under the International Federation for Clinical Chemistry and Laboratory Medicine (IFCC) and International Union for Pure and Applied Chemistry (IUPAC) has developed a systematic nomenclature for the correct classification of laboratory investigations. Each generic laboratory investigation is provided with a unique NPU code. The system is in use among approximately 30 different clinical laboratories in Sweden, and has capacity to be the common denominator of all laboratory investigations, and to be used as the identifier in various information systems. The NPU system for the Swedish laboratories is currently administered by EQUALIS and partly financed by the participating laboratories. Other ways of funding, of benefit for the whole health care sector, will be investigated.

Identification of randomly selected colonies of lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA variable V1 and V3 regions.

The present study aimed to characterize lactobacilli in vaginal fluid from 23 adult healthy women by using high-throughput DNA sequencing for identification of a large number of randomly selected colonies appearing on Rogosa and blood agar. The typing method was based on broad-range PCR of 16S rRNA gene variable regions V1 and V3, pyrosequencing, and classification of the fragments by alignment with NCBI-catalogued sequences and type strain sequences. Four major groups of sequences were found among the 402 isolates clearly corresponding to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii when compared to the sequences obtained for type strains. Our results indicate that pyrosequencing of 16S rRNA gene fragments as used here is a fast and reliable method well suited for identification to the species level, even within the Lactobacillus acidophilus complex.

Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis.

The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297-301). We also diagnosed BV according to Hallén et al. (A. Hallén et al. Genitourin Med 1987;63:386-9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallén et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 microg/g vaginal fluid, do not always correlate with BV.

Terms used to describe urinary tract infections--the importance of conceptual clarification.

Inaccuracies in medical language are detrimental to communication and statistics in medicine, and thereby to clinical practice, medical science and public health. The purpose of this article is to explore inconsistencies in the use of some medical terms: urinary tract infection, bacteriuria and urethral syndrome. The investigated literature was collected from medical dictionaries, textbooks, and articles indexed in Medline(R). We found various practices regarding how the medical terms should be defined, and had great difficulty in interpreting the status of the statements under the heading of 'definition'. The lesson to be learned, besides a reminder of the importance of clearly defined medical concepts, is that it must be explicitly stated whether what is presented as a definition is to be considered as defining criterion, as recognising criterion or as characteristic of the disease entity.

Bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction.

The occurrence of bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction who used clean intermittent catheterisation to empty their bladders was studied in order to examine cut-off concentration breakpoints for significant bacteriuria in this group of patients using procedures of the European Urinanalysis Guideline. 344 samples were cultured, yielding 285 isolates. Coagulase-negative staphylococci (27%), Enterococci (25%), Klebsiella spp (19%), and Escherichia coli (12%) were the most common findings. Bacteria grew at concentrations of 10(5)-10(8) cfu/L, but only a few at 10(4) cfu/L. It is concluded that low bacterial concentrations in the urine (10(5) cfu/L) of patients with neurogenic bladder dysfunction who are on intermittent catheterisation might be as significant for bladder contamination with bacteria as a high bacterial concentration and can possibly be responsible for bladder infections.

Using the MEDLINE database to study the concept of urinary tract infections in different domains of medicine.

As a way of exploring differences between medical domains regarding management of urinary tract infections, we investigated the MEDLINE database for differences in indexing patterns. Further, our intention was to assess the MEDLINE database as a source for studying medical domains. We examined the use of main headings, subheadings and the level of main headings in six medical domains that manage urinary tract infections. Many intuitive but also some counterintuitive results were found indicating that the MEDLINE database is difficult to use for studying medical domains mainly due to unclear semantics both in the headings and the indexing process, which results in variability in indexing. This variability probably hides significant results. We also conclude that the differences found indicate that in addition to differences between domains, there are also large variations within domains.

Vaginal retention of locally administered clindamycin.

Since bacterial vaginosis (BV) is characterized by a lack of, or very few, lactobacilli and high numbers of small, mostly anaerobic bacteria, an obvious treatment modality would be eradication of the BV-associated bacterial flora followed by reintroduction of lactobacilli vaginally. As probiotic treatment with lactobacilli is one tool for improving the cure rate when treating BV, it is necessary to know the length of time after treatment that clindamycin can be found in the vagina and if this could interfere with the growth of the probiotic lactobacilli. We evaluated the vaginal concentration of clindamycin in 12 women for 8 days to obtain data on the concentration of clindamycin in the vagina after intravaginal treatment with the drug. The participants were examined five times between two menstrual periods: before treatment, the day after treatment was finished, and 3, 5 and 8 days post-treatment. The first day post-treatment clindamycin 0.46 × 10(-3) to 8.4 × 10(-3) g/g vaginal fluid (median 2.87 × 10(-3)) was found. Thereafter, the concentration of clindamycin decreased rapidly. In 10 patients clindamycin was found after 3 days. A very low concentration was still present 5 days after treatment in four patients. After 8 days no clindamycin was found. Clindamycin is rapidly eliminated from the vagina, within 3-8 days, after local administration. Our results indicate that treatment with probiotic lactobacilli could be problematic if carried out within 5 days after cessation of clindamycin treatment.