Convergent evolution in high-altitude and marine mammals:Molecular adaptations to pulmonary fibrosis and hypoxia

High-altitude and marine mammals inhabit distinct ecosystems but share a common challenge:hypoxia.To survive in low-oxygen environments,these species have evolved similar phenotypic pulmonary adaptations,characterized by a high density of elastic fibers.In this study,we explored the molecular mechanisms underlying these adaptations,focusing on pulmonary fibrosis and hypoxia tolerance through comparative genomics and convergent evolution analyses.We observed significant expansions and contractions in certain gene families across both high-altitude and marine mammals,closely associated with processes involved in pulmonary fibrosis.Notably,members of the keratin gene family,such as KRT17 and KRT14,appear to be associated with the development of the dense elastic fiber phenotype observed in the lungs of hypoxia-tolerant mammals.Through selection pressure and amino acid substitution analyses,we identified multiple genes exhibiting convergent accelerated evolution,positive selection,and amino acid substitution in these species,associated with adaptation to hypoxic environments.Specifically,the convergent evolution of ZFP36L1,FN1,and NEDD9 was found to contribute to the high density of elastic fibers in the lungs of both high-altitude and marine mammals,facilitating their hypoxia tolerance.Additionally,we identified convergent amino acid substitutions and gene loss events associated with sperm development,differentiation,and spermatogenesis,such as amino acid substitutions in SLC26A3 and pseudogenization of CFAP47,as confirmed by PCR.These genetic alterations may be linked to changes in the reproductive capabilities of these animals.Overall,this study offers novel perspectives on the genetic and molecular adaptations of high-altitude and marine mammals to hypoxic environments,with a particular emphasis on pulmonary fibrosis.

Clinical efficacy of intradermal type Ⅰ collagen injections in treating skin photoaging in patients from high-altitude areas

BACKGROUND Photoaging,a result of chronic sun exposure,leads to skin damage and pigmentation changes.Traditional treatments may have limitations in high-altitude areas like Yunnan Province.Intradermal Col Ⅰ injections stimulate collagen production,potentially improving skin quality.This study aims to assess the efficacy and safety of this treatment for photoaging.AIM To evaluate the efficacy and safety of intradermal typeΙcollagen(ColΙ)injection for treating photoaging.METHODS This prospective,self-controlled study investigated the impact of intradermal injections of ColΙon skin photodamage in 20 patients from the Yunnan Province.Total six treatment sessions were conducted every 4 wk±3 d.Before and after each treatment,facial skin characteristics were quantified using a VISIA skin detector.Skin thickness data were assessed using the ultrasound probes of the Dermalab skin detector.The Face-Q scale was used for subjective evaluation of the treatment effect by the patients.RESULTS The skin thickness of the right cheek consistently increased after each treatment session compared with baseline.The skin thickness of the left cheek significantly increased after the third through sixth treatment sessions compared with baseline.The skin thickness of the right zygomatic region increased after the second to sixth treatment sessions,whereas that of the left zygomatic region showed a significant increase after the fourth through sixth treatment sessions.The skin thickness of both temporal regions significantly increased after the fifth and sixth treatment sessions compared with baseline(P<0.05).These findings were also supported by skin ultrasound images.The feature count for the red areas and wrinkle feature count decreased following the treatment(P<0.05).VISIA assessments also revealed a decrease in the red areas after treatment.The Face-QSatisfaction with Facial Appearance Overall and Face-Q-Satisfaction with Skin scores significantly increased after each treatment session.The overall appearance of the patients improved after treatment.CONCLUSION Intradermal ColΙinjection improves photoaging,with higher patient satisfaction and fewer adverse reactions,and could be an effective treatment method for populations residing in high-altitude areas.

Atmospheric transmission algorithm for pulsed X-rays from high-altitude nuclear detonations based on scattering correction

In high-altitude nuclear detonations,the proportion of pulsed X-ray energy can exceed 70%,making it a specific monitoring signal for such events.These pulsed X-rays can be captured using a satellite-borne X-ray detector following atmospheric transmission.To quantitatively analyze the effects of different satellite detection altitudes,burst heights,and transmission angles on the physical processes of X-ray transport and energy fluence,we developed an atmospheric transmission algorithm for pulsed X-rays from high-altitude nuclear detonations based on scattering correction.The proposed method is an improvement over the traditional analytical method that only computes direct-transmission X-rays.The traditional analytical method exhibits a maximum relative error of 67.79% compared with the Monte Carlo method.Our improved method reduces this error to within 10% under the same conditions,even reaching 1% in certain scenarios.Moreover,its computation time is 48,000 times faster than that of the Monte Carlo method.These results have important theoretical significance and engineering application value for designing satellite-borne nuclear detonation pulsed X-ray detectors,inverting nuclear detonation source terms,and assessing ionospheric effects.

Single-Arm Clinical Study of Combination Perindopril-Amlodipine Tablets in the Treatment of High-Altitude Hypertension

Objective:To evaluate the efficacy of combination perindopril/amlodipine tablets in patients with high-altitude hypertension who were previously unable to control their blood pressure with monotherapy.Methods:A total of 151 patients with high-altitude hypertension whose blood pressure remained inadequately controlled with previous monotherapy were enrolled in this study.All patients received an 8-week treatment with a combination of perindopril/amlodipine tablets,consisting of perindopril 10 mg/day and amlodipine 5 mg/day.Blood pressure measurements,including both diastolic and systolic pressures,were taken at baseline,and after 2,4,6,and 8 weeks of treatment.Results:After 8 weeks of treatment,there was a significant reduction in both average systolic and diastolic blood pressure compared to baseline(P<0.0001).Specifically,the average systolic blood pressure decreased by 24.45±13.75 mmHg,and the average diastolic blood pressure decreased by 13.37±8.40 mmHg.The overall heart rate showed no significant changes during the treatment period.Conclusion:A combination of perindopril/amlodipine tablets significantly improved blood pressure control in patients with high-altitude hypertension after 8 weeks of treatment.These results support the efficacy of combination perindopril/amlodipine as a viable treatment option for high-altitude hypertension.

地黄花叶病毒RT-LAMP快速检测方法的建立及应用

基于反转录环介导等温扩增(reverse transcription loop-mediated isothermal amplification,RT-LAMP)技术建立了针对地黄花叶病毒(Rehmannia mosaic virus,ReMV)的快速、特异、灵敏的RT-LAMP检测方法。根据ReMV外壳蛋白(coat protein,CP)基因的核苷酸序列设计了3组LAMP引物,通过引物筛选确定最佳引物组合;利用电泳检测及可视化方法(加SYBR GreenⅠ显色剂)进行RT-LAMP温度优化、特异性检测、灵敏度比较。结果表明:本研究建立的RT-LAMP方法最适检测温度为60℃,优化后的LAMP方法能特异性检测ReMV,灵敏度是常规PCR的1000倍,可检测到1.2×10^(-1)拷贝/μL的模板浓度。对60份地黄样品的检测结果表明,RT-LAMP方法的阳性检出率为98.3%,高于常规RT-PCR方法(75%)。本研究建立的RT-LAMP检测技术为ReMV的准确检测提供了快速、高效的方法。

Correlation of vascular endothelial growth factor to permeability of blood-brain barrier and brain edema during high-altitude exposure

BACKGROUND： Many studies have evaluated the role of vascular endothelial growth factor （VEGF） in traumatic brain edema and hemorrhagic brain edema. OBJECTIVE： To observe the effects of VEGF expression on permeability of the blood-brain barrier （BBB） during high-altitude and hypoxia exposure, and to investigate the correlation between VEGF expression and BBB permeability with regard to Evans blue staining and brain edema during high-altitude exposure. DESIGN, TIME AND SETTING： The randomized, controlled, animal study was performed at the Tanggula Etape, Central Laboratory of Chengdu Medical College, and Central Laboratory of General Hospital of Chengdu Military Area Command of Chinese PLA, China, from July 2003 to November 2004. MATERIALS： Quantitative RT-PCR kit （Sigma, USA）, VEGF ELISA kit （Biosource, USA）, and Evans blue （Jingchun, China） were acquired for this study. METHODS： A total of 180 Wistar rats were equally and randomly assigned to 15 groups： low-altitude （500 m）, middle-altitude （2 880 m）, high-altitude （4 200 m）, super-high-altitude （5 000 m）, 1,3, 5, 7, 9, 11, 13, 15, 17, 19, and 21 days of super high-altitude exposure. Wistar rats were exposed to various altitude gradients to establish a hypoxia model. MAIN OUTCOME MEASURES： Brain water content was calculated according to the wet-to-dry weight ratio. BBB permeability to Evans blue was determined by colorimetric method. VEGF mRNA and protein levels in brain tissues were detected using RT-PCR and double-antibody sandwich ELISA. RESULTS： Brain water content, BBB permeability to Evans blue, and VEGF mRNA and protein levels in brain tissues increased with increasing altitude and prolonged exposure to altitude. The greatest increase was determined on day 9 upon ascending 5 000 m. Simultaneously, VEGF expression positively correlated to BBB permeability of Evans blue and brain water content （r = 0.975, 0.917, P〈 0.01）. CONCLUSION： Increased VEGF protein and mRNA expression was responsible for increased BBB permeability, which may be an important mechanism underlying brain edema during high-altitude exposure.

Geostructures, dynamics and risk mitigation of high-altitude and long- runout rockslides

Long-runout rockslides at high altitude could cause disaster chain in river basins and destroy towns and major infrasturctures.This paper firstly explores the initiation mechanism of high-altitude and long-runout rockslides.Two types of sliding-prone geostructure models,i.e.the fault control type in orogenic belt and the fold control type in platform area,are proposed.Then,large-scale experimental apparatus and associated numerical simulations are conducted to understanding the chain-style dynamics of rockslide-debris avalanche-debris flow.The results reveal the fragmentation effects,the rheological behaviors and the boundary layer effect of long-runout avalanche-debris flow.The dynamic character-istics of quasi-static-transition-inertia state and solid-liquid coupling in rapid movement of rockslide-debris avalanche-debris flow are investigated.Finally,the risk mitigation strategy of the non-structure and structure for resilient energy dissipation are illustrated for initiation,transition and deposition zones.The structural prevention and mitigation methods have been successfully applied to the high-altitude and long-runout rockslides in Zhouqu and Maoxian of the Wenchuan earthquake zone,as well as the other major geohazards in Qinghai-Tibet Plateau and its adjacent areas.

Comprehensive Marine Observing Experiment Based on High-Altitude Large Unmanned Aerial Vehicle(South China Sea Experiment2020 of the “Petrel Project”)

In collaboration with 12 other institutions, the Meteorological Observation Center of the China Meteorological Administration undertook a comprehensive marine observation experiment in the South China Sea using the Yilong-10 high-altitude large unmanned aerial vehicle(UAV). The Yilong-10 UAV carried a self-developed dropsonde system and a millimeter-wave cloud radar system. In addition, a solar-powered unmanned surface vessel and two drifting buoys were used. The experiment was further supported by an intelligent, reciprocating horizontal drifting radiosonde system that was deployed from the Sansha Meteorological Observing Station, with the intent of producing a stereoscopic observation over the South China Sea. Comprehensive three-dimensional observations were collected using the system from 31 July to2 August, 2020. This information was used to investigate the formation and development processes of Typhoon Sinlaku(2020). The data contain measurements of 21 oceanic and meteorological parameters acquired by the five devices, along with video footage from the UAV. The data proved very helpful in determining the actual location and intensity of Typhoon Sinlaku(2020). The experiment demonstrates the feasibility of using a high-altitude, large UAV to fill in the gaps between operational meteorological observations of marine areas and typhoons near China, and marks a milestone for the use of such data for analyzing the structure and impact of a typhoon in the South China Sea. It also demonstrates the potential for establishing operational UAV meteorological observing systems in the future, and the assimilation of such data into numerical weather prediction models.

Industrial Poverty Alleviation Model in Southwestern High-altitude Mountainous Areas of China——A Case Study of Industrial Poverty Alleviation of Xueshan Township in Luquan County of Yunnan Province through Planting Codonopsis pilosula

Due to the severe restrictions of natural conditions and ecological environment,high-altitude mountainous areas usually become the " hard bones" in the battle against poverty. Xueshan Township,Luquan Yi and Miao Autonomous County of Yunnan Province,located in the alpine valley of Jinsha River,is a major township with wide and deep poverty,and the incidence of poverty is up to 45. 00%. In recent years,Xueshan Township has insisted on the battle against poverty,made effort to develop the Codonopsis pilosula industry,and successfully developed a road to poverty alleviation through C. pilosula industry,and formed a unique industrial poverty alleviation model by the end of 2018,the incidence of poverty dropped to 0. 74%. Based on field survey and interview,this paper analyzes and summarizes the specific practices,main results,practical experience and promotion and application measures of the poverty alleviation model of C. pilosula planting industry in Xueshan Township,in the hope of providing certain reference for the targeted poverty alleviation in similar areas in Yunnan Province and other provinces of China.

Results of an Intentional Introduction of Artemia sinica in the High-Altitude Tibetan Lake Dangxiong Co:On a Base of Surveys in 2011 and 2013

1 Introduction The human population is projected to reach 9200million by 2050.A fundamental question for science is whether it is possible to increase food production to meet the demands of a human population of that magnitude.It’s possible only to increase an aquaculture production.Cultivation of fish/shrimp larvae is a bottleneck in a cultivation of the different organisms.Live food

Exendin-4 inhibits high-altitude cerebral edema by protecting against neurobiological dysfunction

The anti-inflammatory and antioxidant effects of exendin-4（Ex-4） have been reported previously.However,whether（Ex-4） has anti-inflammatory and antioxidant effects on high-altitude cerebral edema（HACE） remains poorly understood.In this study,two rat models of HACE were established by placing rats in a hypoxic environment with a simulated altitude of either 6000-or 7000-m above sea level（MASL） for 72 hours.An altitude of 7000 MASL with 72-hours of hypoxia was found to be the optimized experimental paradigm for establishing HACE models.Then,in rats where a model of HACE was established by introducing them to a 7000 MASL environment with 72-hours of hypoxia treatment,2,10 and,100 μg of Ex-4 was intraperitoneally administrated.The open field test and tail suspension test were used to test animal behavior.Routine methods were used to detect change in inflammatory cells.Hematoxylin-eosin staining was performed to determine pathological changes to brain tissue.Wet/dry weight ratios were used to measure brain water content.Evans blue leakage was used to determine blood-brain barrier integrity.Enzyme-linked immunosorbent assay（ELISA） was performed to measure markers of inflammation and oxidative stress including superoxide dismutase,glutathione,and malonaldehyde values,as well as interleukin-6,tumor necrosis factor-alpha,cyclic adenosine monophosphate levels in the brain tissue.Western blot analysis was performed to determine the levels of occludin,ZO-1,SOCS-3,vascular endothelial growth factor,EPAC1,nuclear factor-kappa B,and aquaporin-4.Our results demonstrate that Ex-4 preconditioning decreased brain water content,inhibited inflammation and oxidative stress,alleviated brain tissue injury,maintain blood-brain barrier integrity,and effectively improved motor function in rat models of HACE.These findings suggest that Ex-4 exhibits therapeutic potential in the treatment of HACE.

鸽新城疫病毒可视化LAMP检测方法的建立与应用

为建立一种简便、快速,适用于基层检测鸽Ⅰ型副黏病毒的方法,研究针对鸽Ⅰ型副黏病毒全基因组保守序列,设计了1组可检测基因Ⅵ、Ⅶ、ⅩⅩ、ⅩⅪ型分离株的环介导等温扩增(LAMP)引物,通过优化反应条件及体系,建立鸽Ⅰ型副黏病毒可视化LAMP检测方法。结果显示:建立的鸽Ⅰ型副黏病毒可视化LAMP检测方法可检测基因Ⅵ、Ⅶ、ⅩⅩ、ⅩⅪ型鸽新城疫病毒,与H9亚型禽流感病毒、马立克病病毒、禽网状内皮组织增殖病病毒、小鹅瘟病毒以及禽白血病病毒以及新城疫病毒La Sota疫苗株均无交叉反应,最低检测限为10^(1)copies/μL。研究表明所建立的鸽Ⅰ型副黏病毒可视化LAMP检测方法快速、简便,特异性强,灵敏度高,可应用于基层快速检测鸽Ⅰ型副黏病毒。

鸡圆圈病毒3型可视化LAMP检测方法的建立及应用

建立鸡圆圈病毒3型(GyV3)可视化环介导等温扩增(LAMP)检测方法,为临床上快速检测鸡GyV3提供一种诊断方法。参考GyV3的VP3基因序列,设计1套LAMP特异性引物,分别通过温度优化、引物浓度优化和反应时间的优化,确定鸡GyV3可视化LAMP诊断方法的扩增温度、引物浓度和反应时间,通过特异性试验、敏感性试验、重复性试验和临床样品检测,验证鸡GyV3可视化LAMP诊断方法的特异性、敏感性、稳定性和可靠性。结果显示,鸡GyV3 LAMP检测方法的反应体系:总体积为20.0μL,包括DNA模板1.0μL,2×Master Mix 10.0μL,内引物GyV3-FIP和GyV3-BIP混合液(工作浓度16.0μmol/L)2.0μL,外引物GyV3-F3和GyV3-B3混合液(工作浓度2.0μmol/L)2.0μL,环引物GyV3-LF和GyV3-LB混合液(工作浓度8.0μmol/L)2.0μL,ddH_(2)O 3μL;鸡GyV3 LAMP检测方法的扩增程序:66℃反应20 min,80℃灭活5 min。鸡GyV3可视化LAMP检测方法能特异性检测GyV3,最低检测浓度为10^(1)拷贝/μL,组内和组间变异系数小于5%;与常规PCR方法检测临床样品的结果比较,两者结果符合率达100%。建立的鸡GyV3可视化LAMP检测方法能特异性检测GyV3,具有特异性好、敏感性高及污染小等优点,且检测结果可通过肉眼观察进行判断,适用于GyV3的临床快速筛查。

基于LAMP技术的欧洲樱桃绕实蝇检测方法的建立

在机场、海港和陆运口岸等入境场所建立快速的现场检疫鉴定方法能够有效防止外来有害生物通过全球商品贸易、旅客携带等方式进行传播和扩散,从而避免造成严重的经济损失。本研究根据欧洲樱桃绕实蝇线粒体细胞色素COI基因序列设计特异的LAMP引物,分别研制了可视化LAMP检测方法和实时荧光LAMP检测方法。结果表明,优化后的可视化LAMP检测体系在62℃、40 min内可检测低至10pg/μL的欧洲樱桃绕实蝇基因组DNA,且不与其他近似种样品发生交叉反应,可以裸眼直接判断检测结果,具有好的特异性;实时荧光LAMP检测方法的最低检测限为10 ng/μL,检测时间约需90 min。本研究建立的快速、准确、灵敏的可视化恒温扩增体系,可以应用于欧洲樱桃绕实蝇的现场快速检测。

Multiyear Observations of Deposition-Mode Ice Nucleating Particles at Two High-Altitude Stations in India

Ice nucleating particle(INP) measurements were made at two high-altitude stations in India. Aerosols collected on filter paper at Girawali Observatory, Inter University Center for Astronomy & Astrophysics(IGO), and at the Radio Astronomy Center, Ooty(RAC), were activated in deposition mode using a thermal gradient diffusion chamber to determine the INP concentrations. The measurement campaigns at IGO were conducted during 2011, 2013 and 2014, and at RAC during 2013 and 2014. When the aerosol samples were exposed to an ice supersaturation of between 5% and 23% in the temperature range-17.6?C to-22?C, the maximum INP number concentration at IGO and RAC was 1.0 L-1and 1.6 L-1, respectively.A maximum correlation coefficient of 0.76 was observed between the INP number concentration and ice supersaturation. The airmass trajectories analyzed for the measurement campaigns showed that the Arabian Desert and arid regions were the main INP contributors. Elemental analysis of particles showed the presence of Na, Cl, Si, Al, Fe, Cu, Co, Cd, S, Mn and K, as well as some rare-Earth elements like Mo, Ru, La, Ce, V and Zr. When aerosols in the size range 0.5–20 μm were considered, the fraction that acted as INPs was 1 : 104to 1 : 106at IGO, and 1 : 103to 1 : 104at RAC. The higher ratio of INPs to aerosols at RAC than IGO may be attributable to the presence of rare-Earth elements observed in the aerosol samples at RAC, which were absent at IGO.

Minimally invasive surgery vs laparotomy in patients with colon cancer residing in high-altitude areas

BACKGROUND Colon cancer is associated with a higher incidence among residents in highaltitude areas.Hypoxic environment at high altitudes inhibits the phagocytic and oxygen-dependent killing function of phagocytes,thereby increasing the inflammatory factors,inhibiting the body’s innate immunity and increasing the risk of colon cancer.AIM To examine the effect of minimally invasive surgery vs laparotomy in patients with colon cancer residing in high-altitude areas.METHODS Ninety-two patients with colon cancer in our hospital from January 2019 to February 2021 were selected and divided into the minimally invasive surgery and laparotomy groups using the random number table method,with 46 patients in each group.Minimally invasive surgery was performed in the minimally invasive group and laparotomy in the laparotomy group.Operative conditions,inflammatory index pre-and post-surgery,immune function index and complication probability were measured.RESULTS Operative duration was significantly longer and intraoperative blood loss and recovery time of gastrointestinal function were significantly less(all P<0.05)in the minimally invasive group than in the laparotomy group.The number of lymph nodes dissected was not significantly different.Before surgery,there were no significant differences in serum C-reactive protein,interleukin-6 and tumor necrosis factor-αlevels between the groups,whereas after surgery,the levels were significantly higher in the minimally invasive group(26.98±6.91 mg/L,146.38±11.23 ng/mL and 83.51±8.69 pg/mL vs 41.15±8.39 mg/L,186.79±15.36 ng/mL and 110.65±12.84 pg/mL,respectively,P<0.05).Furthermore,before surgery,there were no significant differences in CD3+,CD4+and CD4+/CD8+levels between the groups,whereas after surgery,the levels decreased in both groups,being significantly higher in the minimally invasive group(55.61%±4.39%,35.45%±3.67%and 1.30±0.35 vs 49.68%±5.33%,31.21%±3.25%and 1.13±0.30,respectively,P<0.05).Complication probability was significantly lower in the minimally invasive group(4.35%vs 17.39%,P<0.05).CONCLUSION Laparoscopic minimally invasive procedures reduce surgical trauma and alleviate the inflammatory response and immune dysfunction caused by invasive operation.It also shortens recovery time and reduces complication probability.

草莓白化伴随病毒RT-LAMP检测方法的建立

草莓白化伴随病毒(strawberry pallidosis-associated virus,SPaV)能引起草莓白化病,与其他病毒复合侵染后会加重症状。本研究建立了SPaV的反转录环介导等温扩增方法(reverse transcription-loop-mediated isothermal amplification,RT-LAMP),对反应体系的各条件进行评估优化后,检测了RT-LAMP的特异性和灵敏度,且对田间样品进行了测试。结果表明,优化后的反应体系包含2.5μL 10×Isothermal Amplification Buffer、6 mmol/L Mg^(2+)、1 mmol/L dNTPs、1μmol/L SPaV-FIP和SPaV-BIP,0.1μmol/L SPaV-F3和SPaV-B3,0.3μmol/L SPaV-LB,0.4 mol/L Betaine,0.256 U/μL WarmStart Bst 2.0 DNA Polymerase,0.12 U/μL WarmStart RTx Reverse Transcriptase,1μL RNA,反应条件为61℃恒温孵育40 min。采用优化后的反应体系,在特异性检测中,只有携带了病毒SPaV的样品才能发生阳性扩增。灵敏度测试中,RT-LAMP比RT-PCR灵敏度高10倍。田间应用中,RT-LAMP和RT-PCR检测结果一致。本研究建立的SPaV RT-LAMP检测方法操作简便、快速、特异,可供实验室及生产实践中应用,助力脱毒种苗的鉴定和田间病毒病调查。

Study on Query System of the Historical High-altitude Ground Weather Chart

We introduced query system of the historical high-altitude surface weather chart. Historical high-altitude surface data were converted to Grads data format. Grads as generation tool of the image, ASP was used to compile WEB page. By B/S mode, only user submitted necessary conditions for the image to server by client browser, historical high-altitude surface weather chart at corresponding time and height could be ob- tained. Without any procedures and related data, only needed client browser, user could use image conveniently to a large extent.

鳞球茎茎线虫LAMP检测方法的建立

鳞球茎茎线虫(Ditylenchus dipsaci)能引起多种植物毁灭性病害,是世界公认的危险性线虫,被列入我国检疫性有害生物名录。本研究通过比较GenBank数据库中茎属线虫相关序列,以鳞球茎茎线虫rDNA-ITS序列为靶标,设计鳞球茎茎线虫LAMP特异性引物,建立了可视化LAMP检测体系。结果表明,本研究建立的LAMP检测方法可在1 h内对鳞球茎茎线虫进行检测,供试其他22种植物线虫均未出现扩增,仅鳞球茎茎线虫出现扩增,具有良好的特异性;检测灵敏度可达1/4000条鳞球茎茎线虫;检测结果可以直接用肉眼判断。因此,本方法具有反应时间短、灵敏度高、设备要求低的优点,适于基层实验室开展鳞球茎茎线虫检测。

虾类十足目虹彩病毒1(DIV1)LAMP-HNB快检方法的建立

[目的]建立一种操作简便、快速、可视化的虾类病原检测方法,实现基层养殖户自检。[方法]针对十足目虹彩病毒1(DIV1),设计环介导等温扩增(LAMP)特异性引物,对反应试剂用量、温度、反应时间等条件进行优化,建立LAMP与羟基萘酚蓝(HNB)结合的LAMP-HNB可视化快速检测方法。[结果]在LAMP反应体系中添加12 mmol/L Mg^(2+)和250μmol/L HNB,65℃恒温条件下反应60 min最佳;DIV1病原质粒最低检出限为102 copies/μL,稳定性和特异性较好,通过带有靶标基因的阳性质粒进行16次重复性试验结果均较为稳定,多种模板扩增试验均未发生假阳性情况,符合试验要求。[结论]该研究建立的LAMP-HNB可视化快速检测方法,适用于上述对虾DIV1的现场快速检测。