A rapid seminested polymerase chain reaction (PCR) method for the specific, sensitive detection of virulent Shigella spp. in spiked environmental water samples was developed. A set of primers specific for the invasion plasmid antigen gene (ipaH) of virulent Shigella spp. and enteroinvasive Escherichia coli produced a 620-bp fragment that was used as template for the seminested primer pair delin...

The presence of PCR inhibitors in water samples is well known and contributes to the fact that a practical PCR assay has not been developed for legionella surveillance. In this study, we devised a new seminested PCR assay for detection of Legionella spp. in water samples as a means of overriding the PCR inhibitors without loss of sensitivity. The seminested PCR assay utilized primers to amplify...

OBJECTIVE The aim of this study was to determine whether or not a noninvasive procedure utilizing maternal peripheral blood as the source of DNA and polymerase chain reaction (PCR) could be used to detect fetal rhesus D (RhD) status as well as fetal gender during different gestational stages of pregnancy. MATERIALS AND METHODS Maternal blood samples were obtained from 54 RhD-negative pregnant...

There is a growing appreciation of the potential value for routine screening for the presence of HPV not only for cervical specimens but also from oral cavity. The purpose of this study was to develop and clinically evaluate a single-tube seminested PCR assay for the detection of HPV. Several parameters such as PCR primers, primer annealing temperature, the number of PCR cycles and concentratio...

A prospective study (from August 2006 to April 2007) was carried out with 214 cerebrospinal fluid samples with suspicion of bacterial meningitis. The aim of the study was to assess the effectiveness of the simultaneous detection of Neisseria meningitidis, Streptococcus sp. and Haemophilus influenzae using seminested polymerase chain reaction strategy. Among the 214 samples tested by both PCR an...

Cell-associated (CA) HIV-1 RNA is considered a potential marker for assessment of viral reservoir dynamics and antiretroviral therapy (ART) response in HIV-infected patients. Recent studies employed sensitive seminested real-time quantitative (q)PCR to quantify CA HIV-1 RNA. Digital PCR has been recently described as an alternative PCR-based technique for absolute quantification with higher acc...

Specific and sensitive reverse transcription-PCR (RT-PCR) assays were developed for the detection of eastern, western, and Venezuelan equine encephalitis viruses (EEE, WEE, and VEE, respectively). Tests for specificity included all known alphavirus species. The EEE-specific RT-PCR amplified a 464-bp region of the E2 gene exclusively from 10 different EEE strains from South and North America wit...

The sequencing of the VP1 hypervariable region of the human enterovirus (HEV) genome has become the reference test for typing field isolates. This study describes a new strategy for typing HEV at the serotype level that uses a reverse transcription-PCR assay targeting the central part of the VP2 capsid protein. Two pairs of primers were used to amplify a fragment of 584 bp (with reference to th...

| An epidemiological study of babesiosis in stray dogs was conducted at Chittagong Metropolitan area, Bangladesh, for six months. Blood samples from 130 stray dogs were collected from nine randomly selected areas along with the information of age and sex. Blood samples were initially examined by Giemsa’s stained blood smear method and DNA was extracted and Babesia spp. were confirmed by amplify...

PCR is the best method for the detection of enteric viruses present at low concentrations in environmental samples. However, some organic and inorganic compounds present in these samples can interfere in the reaction. Many of these substances are cytotoxic, too. The ZP60S filter membranes used in addition to fluorpentane treatment are quite efficient for virus concentration and simultaneous eli...