New molecular target for the phylogenetic identification of Leptospira species directly from clinical samples: an alternative gene to 16S rRNA

Abstract INTRODUCTION: Phylogenetic analysis of the 16S ribosomal gene initial region is used to identify Leptospira isolates at the species level from clinical samples. Unfortunately, this method cannot differentiate between some intermediates and saprophytic species. METHODS: We used comparative genomic analysis between 35 Leptospira species to find new molecular targets for Leptospira species identification. RESULTS: We proposed the use of the rpoC gene, encoding the DNA-directed RNA polymerase β-subunit, for identifying 35 Leptospira species. CONCLUSIONS: The rpoC gene can be a molecular target to identify the main species of the Leptospira genus directly from clinical samples.

Hypervascular retained products of conception: dilemma of diagnosis and management

Retained products of conception (RPOC) are a partial retention of placental tissue after dilation and curettage (D&C) procedures or vaginal deliveries. Ultrasound scan reports sometimes mention the presence of increased endometrial / sub‐endometrial vascularity in the context of retained products of conception. This raises the possibility of serious intra‐operative haemorrhage because of the possibility of arterio‐venous malformation. The aim of this article is to discuss the diagnosis and management options of retained products of conception (RPOC) with increased vascularity where simple dilatation and curettage may lead to life threatening haemorrhage and endanger the life of the patient and to enlighten the importance of evaluation of vascularity in all cases of RPOC prior to dilatation and curettage in order to avoid the dreaded complication of massive haemorrhage.

Identification and genetic diversity analysis of Memecylon species using ISSR, RAPD and Gene-Based DNA barcoding tools

Background: Memecylon species are commonly used in Indian ethnomedical practices. The accurate identification is vital to enhance the drug's efficacy and biosafety. In the present study, PCR based techniques like RAPD, ISSR and DNA barcoding regions, such as 5s, psbA-trnH, rpoC1, ndh and atpF-atpH, were used to authenticate and analyze the diversity of five Memecylon species collected from Western Ghats of India. Results: Phylogenetic analysis clearly distinguished Memecylon malabaricum from Memecylon wightii and Memecylon umbellatum from Memecylon edule and clades formed are in accordance with morphological keys. In the RAPD and ISSR analyses, 27 accessions representing five Memecylon species were distinctly separated into three different clades. M. malabaricum and M. wightii grouped together and M. umbellatum, M. edule and Memecylon talbotianum grouped in the same clade with high Jaccard dissimilarity coefficient and bootstrap support between each node, indicating that these grouped species are phylogenetically similar. Conclusion: Data from the present study reveals that chloroplast psbA-trnH region could be used as a potential candidate region for identifying Memecylon species, and ISSR marker system could be used for estimating genetic diversity since it has high percent polymorphism compared to RAPD marker.

Authentication of Angelica sinensis and adulterants by analysis of plastid trnL-F and rpoC1 sequences / 中国药学杂志

OBJECTIVE: To establish a new method for identification of Angelica sinensis from its adulterants by analyzing trnL-F and rpoC1 sequences. METHODS: The plastid trnL-F and rpoC1 of 25 samples of A. sinensis and its adulterants were amplified, sequenced, and analyzed. The K-2-P distances of A. sinensis and its adulterants were calculated, and the phylogenetic trees were constructed. RESULTS: The trnL-F sequence showed significantly larger length variation range, numbers of variable sites and informative sites, and evolutionary distance than rpoC1 sequence. There were significant differences between A. sinensis and the adulterants based on the trnL-F sequences. One SNP site and one repeated base A region could be used as specific authenticable sites. The distance of A. sinensis and its adulterants ranged from 0.002-0.231 as shown by trnL-F sequences. Phylogeny tree reconstruction using MP analysis based on trnL-F sequences could effectively distinguish A. sinensis from adulterants. However, analysis of the rpoC1 sequences could not identify A. sinensis and its adulterants. CONCLUSION: The rpoC1 sequence has poor capability for identification of A. sinensis and its adulterants. The trnL-F sequence could be used as an efficient molecular marker for authenticating A. sinensis from its adulterants.