Subject(s)
Proteome/analysis , Proteomics/methods , Severe Acute Respiratory Syndrome/diagnosis , Viral Proteins/blood , Adult , Aged , Aged, 80 and over , Algorithms , Biomarkers/blood , Case-Control Studies , Female , Humans , Male , Middle Aged , Models, Biological , Prognosis , Protein Array Analysis , Severe Acute Respiratory Syndrome/metabolism , Severe Acute Respiratory Syndrome/virology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Young AdultABSTRACT
Male-specific coliphages are often used as indicators of contamination by enteric viruses. These phages can be detected in water samples by plaque assays and by polymerase chain reaction. In this study, the M13 coliphage was used to develop a real-time PCR assay for the detection of male-specific DNA coliphages. The real-time PCR was found to have a reaction efficiency of 1.45 and detection limit of 10(-3) plaque forming units per reaction mix. Repeated amplification and melting curve analyses demonstrated high specificity and reproducibility of the real-time assay. Quantitative detection with the real-time PCR should allow rapid assessment of the level of viral contamination in water.
Subject(s)
Bacteriophage M13/genetics , Bacteriophage M13/isolation & purification , DNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , Water Microbiology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: It has been postulated that genetic predisposition may influence the susceptibility to SARS-coronavirus infection and disease outcomes. A recent study has suggested that the deletion allele (D allele) of the angiotensin converting enzyme (ACE) gene is associated with hypoxemia in SARS patients. Moreover, the ACE D allele has been shown to be more prevalent in patients suffering from adult respiratory distress syndrome (ARDS) in a previous study. Thus, we have investigated the association between ACE insertion/deletion (I/D) polymorphism and the progression to ARDS or requirement of intensive care in SARS patients. METHOD: One hundred and forty genetically unrelated Chinese SARS patients and 326 healthy volunteers were recruited. The ACE I/D genotypes were determined by polymerase chain reaction and agarose gel electrophoresis. RESULTS: There is no significant difference in the genotypic distributions and the allelic frequencies of the ACE I/D polymorphism between the SARS patients and the healthy control subjects. Moreover, there is also no evidence that ACE I/D polymorphism is associated with the progression to ARDS or the requirement of intensive care in the SARS patients. In multivariate logistic analysis, age is the only factor associated with the development of ARDS while age and male sex are independent factors associated with the requirement of intensive care. CONCLUSION: The ACE I/D polymorphism is not directly related to increased susceptibility to SARS-coronavirus infection and is not associated with poor outcomes after SARS-coronavirus infection.
Subject(s)
Peptidyl-Dipeptidase A/genetics , Polymorphism, Genetic/genetics , Respiratory Distress Syndrome/complications , Respiratory Distress Syndrome/genetics , Severe Acute Respiratory Syndrome/complications , Adult , Aged , Aged, 80 and over , Alleles , Case-Control Studies , Female , Genetic Predisposition to Disease/genetics , Genotype , Humans , Male , Middle Aged , Peptidyl-Dipeptidase A/metabolismSubject(s)
Oligonucleotide Array Sequence Analysis/methods , Placenta/metabolism , RNA, Messenger/metabolism , Female , Gene Expression Profiling , Humans , Pregnancy , Pregnancy Trimester, First/blood , Pregnancy Trimester, First/genetics , Pregnancy Trimester, Third/blood , Pregnancy Trimester, Third/genetics , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction/methodsABSTRACT
Severe acute respiratory syndrome (SARS) is a global health concern. In Hong Kong, two major outbreaks, one hospital based and the other in the Amoy Gardens apartments, were identified. The frequency of diarrhoea, admission to intensive care, and mortality differed significantly between the two outbreaks. We did genomic sequencing for viral isolates from five Amoy Gardens patients. The virus sequence was identical in four of these five patients. The sequence data from one hospital case and the four identical community cases had only three nucleotide differences. Alterations in the SARS coronavirus genome are unlikely to have caused the distinctive clinical features of the Amoy Gardens patients, and these results highlight the importance of non-viral genomic factors in this outbreak.
Subject(s)
Disease Outbreaks/statistics & numerical data , Genome, Viral , Severe Acute Respiratory Syndrome/virology , Severe acute respiratory syndrome-related coronavirus/genetics , Base Sequence/genetics , Cross Infection/virology , Hong Kong/epidemiology , Humans , Severe Acute Respiratory Syndrome/diagnosis , Severe Acute Respiratory Syndrome/epidemiologyABSTRACT
Fifty-two patients with solid tumors had depressed white blood cell and neutrophil counts because of prior treatment with cytotoxic cancer chemotherapeutic drugs. These patients were given one or more i.v. infusions of 0.125-0.25 mg of 12-O-tetradecanoylphorbol-13-acetate (TPA), and this treatment increased the low white blood cell and neutrophil counts toward the normal range. The average white blood cell and neutrophil counts were 2.55 x 10(9)/liter and 1.76 x 10(9)/liter, respectively, before treatment with TPA. After one or more i.v. infusions of TPA, the white blood cell and neutrophil counts increased to peak values of 5. 92 x 10(9)/liter and 4.76 x 10(9)/liter, respectively, within a few days. Most patients had increased levels of white blood cells and neutrophils by 24 hr after a single i.v. infusion of 0.25 mg TPA. Elevated levels were observed for at least 3 days. This study demonstrates that treatment with parenteral TPA is feasible with useful biological activity. Only mild and reversible side effects were observed.
