Establishment of platelet donor gene bank involving specific antigen HPA 1-6/10/15/21 by real-time fluorescent PCR / 中国输血杂志

【Objective】 To research the genetic polymorphism of HPA 1-6/10/15/21 in platelet donors in Zhongshan area, and establish a gene bank of platelet donors with HPA locus. 【Methods】 The HPA 1-6/10/15/21 system genotyping was performed by Real time fluorescent PCR combined with TaqMan probe technology on 192 platelet donors in Zhongshan area, and the genotype frequency and gene frequency were calculated. 【Results】 Only HPA-aa genotype was found within HPA-4/10, and no allele HPA-b had been detected. The majority of HPA-1, 2, 5, 6 and 21 genotypes were aa. HPA-3 and HPA-15 showed high heterozygosity, with genotype frequency of 0.307 3, 0.494 8 and 0.197 9 for HPA- 3aa, HPA-3ab and HPA-3bb, while 0.270 8, 0.505 2 and, 0.224 0 for HPA -15aa, HPA-15ab and HPA-15bb, respectively. 【Conclusion】 The distribution characteristics of HPA 1-6 /10/15/21 of platelet donors in Zhongshan shows regional differences compared with similar researches from other regions. The establishment of HPA gene bank is helpful to avoid alloimmunization caused by incompatible platelet transfusion.

Phylogenetic relationship analysis of common medicinal submetals and multiple real-time fluorescence PCR identification of Fritillariae Cirrhosae Bulbus / 中草药

Objective To investigate the feasibility of multiple real-time PCR for the detection of Fritillariae Cirrhosae Bulbus and adulterants. Methods Based on the analysis of interspecies variation, genetic distance and phylogenetic relationship of ITS, psbA-trnH, rbcL and matK gene sequences, the genes with fast evolution rate, big interspecies variation and small intraspecies variation were selected as target genes. Fritillariae Cirrhosae Bulbus and adulterants specific primers and Taqman probes were designed to establish a multiplex real-time PCR assay. Methods were evaluated by comparison of specificity, sensitivity and mixed sample detection and sequencing. Results The ITS and psbA-trnH mutations were higher than rbcL and matK, and rbcL and matK were significantly lower than ITS and psbA-trnH genes by genetic distance analysis. And the sensitivity of the establish multiple real-time PCR using ITS as the target gene was 0.01 ng. Four samples of adulterants were detected in 18 samples, and the results were consistent with the results of NJ tree clustering analysis. Conclusion Based on the IIS region sequence as the target gene to establish multiple real-time fluorescence PCR detection method can successfully identify Fritillariae Cirrhosae Bulbus and its counterfeit goods, which provides a new basis for the authenticity of identification.

Identification of Ophiocordyceps sinensis and its adulterants based on portable and CFX96 real-time fluorescent PCR systems / 药学学报

The molecular identification of Ophiocordyceps sinensis and its adulterants was carried out by real-time fluorescent PCR with TaqMan probe. Genomic DNA was extracted from 100 samples of Ophiocordyceps sinensis and its adulterants. MEGA 7.0 software was used for comparative analysis to define the variable sites between Ophiocordyceps sinensis and its adulterants according to the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). A set of specific primers and TaqMan probe were designed using Primer Premier 6.0 software, and sensitivity and specificity studies were performed on two different real-time fluorescent PCR systems (Genesig q16 and Bio-Rad CFX96). The sensitivity study showed that the detectable DNA template concentration of Ophiocordyceps sinensis for the real-time fluorescent PCR was 0.016 ng·μL-1 in the Bio-Rad CFX96 system and 15.527 ng·μL-1 in the Genesig q16 system, respectively. Meanwhile, this method had good specificity for Ophiocordyceps sinensis on Genesig q16 and Bio-Rad CFX96 systems, so Ophiocordyceps sinensis could be clearly distinguished from Ophiocordyceps nutans, Cordyceps gunnii, Cordyceps militaris, Cordyceps cicadae, Cordyceps liangshanensis, Cordyceps gracilis. Our results indicate that real-time fluorescent PCR with TaqMan probe can be used to accurately identify Ophiocordyceps sinensis from its adulterants. This provides a technical method that has wide applications for market management and quality control of Chinese materia medica.

Differences of Nuc-mecA Gene Carriage in the Noses among Different Altitudes in High Humidity District / 现代检验医学杂志

Objective To investigate the carriage of nuc-mecA gene among different altitudes in high humidity district,provid-ed guiding data for prevention of staphylococcus aureus and drug-resistant bacteria,standardizing the usage for antibiotics. Methods The nose swabs were collected in different altitudes:1 000 m,1 200 m and 1 400 m,nuc-mecAgene was confirmed by multi-channel real-time PCR.Results The carrier of nuc gene in the noses were 4.878%,2.899% and 7.143%,in 1 000 m,1 200 m and 1 400 m respectively,and there were no statistical significant among the altitudes (P>0.05).The carrier of mecA gene were 14.634%,31.884% and 41.837% in the 1 000 m,1 200 m and 1 400 m respectively,the difference showed statistical significe (P0.05).Conclusion The carrier of mecA gene in noses was increased with the increasing of the altitude. The residents who living at higher altitude should keep the colonization sites of pathogens clean,and needed timely medical when got sick,shouldn’t abuse the antibiotics without authorization.Medical staff should rational use of antibiotic drugs,a-voided overusing of antibiotics and overtreatment.

Analysis of 75 cases of suspected type A influenza results by real-time fluorescent quantitative PCR test / 国际检验医学杂志

Objective To analyze the detection results of 75 suspected type A influenza cases and to understand the distribution situation of influenza patients to provide the guidance for its early prevention and control .Methods The nasopharyngeal swabs specimens from 75 patients with suspected type A influenza were detected for understanding the viral infection situation by real‐time fluorescent quantitative PCR .Results In the specimens of 75 suspected patients ,13 cases with type A influenza virus were detected out ,in which 9 cases were the H1N1 subtype ,mean age > 50 years old .The temporal distribution was 3 cases in March 2013 ,1 case in April ,7 cases in January 2014 and 2 cases in February .No H7N9 subtype was detected out .Conclusion Real‐time fluores‐cent quantitative PCR has the characteristics of rapidness and high accuracy .The winter is the high onset season of flu ,elderly pa‐tients are susceptible to influenza virus .The monitoring and early treatment should be strengthened .

Detection and Identification of Tilletia controversa K?hn by Real-time fluorescent PCR / 微生物学通报

By analysing the ribosome gene(rDNA) sequences in externally transcribed spacer(ETS) of wheat T.controversa(TCK) and its similar spacies T.caries(TCT)and T.foetida(TFL).The special sequences of TCK's ETS have been found,.And designed Taqman probe according to the special sequences,TCK has been successfully detected by using Real-time Fluorescent PCR.