Video在线视频

早上好
-Good morning.

我是Catherine Pierre-Audigier
I am Catherine Pierre-Audigier,

我在巴黎巴斯德研究院的
I work in the Mycobacterial Genetics Unit

分枝杆菌遗传学研究组
at the Institut Pasteur, Paris,

和Bichat-Claude Bernard医院的
and in the clinical microbiology laboratory

临床微生物学实验室工作
in the hospital Bichat-Claude Bernard.

我们将要谈论培养 Hain检测
We are going to speak about cultures, Hain,

以及异烟肼（isoniazid, Inh）
isoniazid

和利福平（rifampicin, RIF)）耐药性
and rifampicin resistance.

对于Luc和Sylvia来说是坏消息
Bad news for Luc and Sylvia,

Xpert结果显示
Xpert results have shown

他们的TB菌株是耐RIF的
that their TB strain is rifampicin-resistant.

而95％的RIF耐药菌株
And 95% of rifampicin-resistant strains

是耐多药((Multidrug resistance, MDR)的
are MDR-TB.

我们怎么知道他们是MDR菌株
How can we know if they are MDR strains?

MTBDRplus Hain试验可确认RIF耐药性
The MTBDRplus Hain test confirms RIF-resistance,

检测Inh耐药性
detects Inh resistance

并且检测临床样品具备足够灵敏度
and is sensitive enough to be performed on clinical samples.

它是如何工作的
How does it work?

用生物素标记的引物扩增目标基因
Biotin label primers amplify the target gene.

特异性探针结合在带膜上
Specific probes are bound on the strip membrane.

检测在TwinCubator上进行
The test is performed on a TwinCubator.

如果扩增产物能识别条带上的探针
If the amplified product recognizes

并与其杂交
and hybridizes the probes on the strip,

染色反应之后可以在条带上显示紫色线
it can be revealed with a purple line on the strip after the staining reaction.

整个试验可以在不到一天的时间内完成
The whole test can be performed in less than a day.

Hain试验证实RIF耐药性
Hain confirms Rif resistance.

rpoB基因的某小段区域的突变
By analyzing a short region in the rpoB gene,

与RIF耐药相关
where mutations are associated with RIF resistance,

Hain试验针对该区域使用八个野生型探针
a Hain test uses eight wild probes all along the region

和四个突变型探针
and four mutated probes

这些探针与最频繁的突变相关
that are associated with the most frequent mutations

并且可以显示为紫色线条
and can be revealed as purple lines on the strip.

那么Inh呢
And what about isoniazid?

Hain试验检测KatG突变
Hain tests detect KatG mutations.

KatG编码的过氧化氢酶
KatG codes a catalase

可以将无活性Inh转化成活化形式
that transforms inactive Inh into the active compound.

KatG基因突变
And mutations in the KatG gene are associated

与Inh高度耐药性有关
with high-level isoniazid resistance.

活化形式的Inh
The active compounds will inhibit InhA,

能抑制分枝菌酸合成中的还原酶InhA
a reductase in the mycolic acid synthesis.

InhA基因突变
Mutations on the InhA gene are associated

与低水平的Inh耐药有关
with low-level Inh resistance.

Luc和Sylvia呢
What about Luc and Sylvia?

Luc感染的是耐RIF菌株
Luc has a RIF-resistant strain.

野生型探针都是阴性的
No wild-type probes are positive

但MUT3是阳性的
but MUT3 is positive,

这标志着识别出了
a signal that recognizes S531L mutations

与RIF耐药高度相关的S531L突变
that are frequently associated with RIF resistance.

而且 对于Inh 野生型探针均为阴性
And also,for Inh,wild-type probes are absent.

MUT1探针阳性
The MUT1 probe is present,

意味着这是一个耐Inh的菌株
that means an Inh-R strain,

也就是耐多药结核病
that means MDR-TB.

那么Sylvia呢
And what about Sylvia?

Syliva感染的是耐RIF菌株
Sylvia has a rifampicin-resistant strain and also,

并且有两个突变
two mutations,

一个在KatG上
one on KatG,

一个在InhA上
one on InhA,

这意味着Inh耐药
that means Inh resistance

并且是耐多药菌株
and an MDR strain.

我们可以对他们使用MDR的治疗方案吗
Can we give MDR treatment?

他们对于二线药物的敏感性如何呢
What about second-line drug susceptibility?

Luc和Sylvia是否患有
Do Luc and Sylvia

广泛耐药肺结核(Extensive Drug Resistant TB, XDR-TB）
have XDR-TB?

为了回答这个问题
To answer the question,

进行MTBDRsl试验分析gyrA
the MTBDRsl test analyzes gyrA

和gyrB基因以检测对喹诺酮类药物的敏感性
and gyrB to test quinolones

分析rrs和eis基因以检测卡那霉素
and the rrs and eis genes to test for kanamycin,

阿米卡星
amikacin capreomycin,

卷曲霉素和紫霉素类抗生素的易感性
viomycin antibiotics susceptibility.

对于Luc
For Luc.

他感染菌株的这些基因的没有突变
Luc has no mutations on these genes.

他感染的菌株对氟喹诺酮类
His strain is sensitive to fluoroquinolones aminoglycosides

氨基糖苷类和卷曲霉素敏感
and capreomycin.

Luc未患有 XDR-TB
Luc does not have XDR-TB.

他可以接受为期九个月的MDR治疗
He can receive a nine-month MDR treatment.

但Sylvia感染的菌株有gyrA突变
But Sylvia,however,

对氟喹诺酮类药物有耐药性
is resistant to fluoroquinolones with a mutation

同时有rrs基因突变
on gyrA and also to aminoglycosides

对氨基糖苷类药物也有耐药性
with a mutation on the rrs gene.

Sylvia患有XDR-TB
Sylvia has XDR-TB.

我们不能对Sylvia 使用MDR标准化治疗方案
We cannot give a standard MDR treatment.

要设计一个最佳的治疗方案
To design an optimal treatment,

我们需要知道什么药物
we need to know what drug can be used to test

可以用来测试哪种药物有效
which drug can be effective.

如何进行药敏试验
How to perform drug susceptibility testing?

一种可提高诊断灵敏度和确定诊断
A way to improve diagnosis sensitivity

然后检测药物敏感性的方法
and to confirm diagnosis and after that, to test drug susceptibility,

是在造价昂贵的生物安全三级实验室
are cultures in a biosafe but expensive laboratory,

开展的菌培
level three.

结核分枝杆菌三个星期内
Mtb grows in three weeks on a solid medium,

在如Ogawa Kudoh或L?wenstein Jensen
such as Ogawa,in the Kudoh method,

培养基的固体培养基上
or L?wenstein Jensen media,

生长成特征性的粗糙菌落
in characteristic rough colonies.

但培养阳性报告晚于临床诊断
But the positive culture is later after the clinical diagnosis.

提高效率和速度的一种方法
A way to improve the efficiency and to be faster

是在液体培养基
is to culture in a liquid medium,

—MGIT培养基中培养
a MGIT medium,

该方法会使Mtb在显微镜下
that gives cores characteristic of Mtb

呈现其核心特征
in microscopy.

此外 进行表型药敏试验
Also, phenotypic drug susceptibility testing,

如果采用液体培养基
either with a liquid medium,

约10天可出结果
with results in about 10 days,

或采用比例法在固体培养基
or in a solid medium, using the proportion method.

三到六周能得到结果
Results will be available in three to six weeks.

总之 分子生物检测试验
In conclusion, molecular tests detect MDR

能够在数小时内检测出MDR和XDR-TB
and XDR-TB within hours.

之后以培养方法进行的表型试验
And later, phenotypic tests

以确认分子生物检测结果
with cultures confirm the molecular results.

以培养方法进行的表型试验耗时较长
They take longer, allowing the patient

但会让病人得到最适当的抗生素治疗
to receive the appropriate antibiotic treatment.

感谢您的关注
Thank you for your attention.